Patent Publication Number: US-2022213226-A1

Title: Anti-tl1a/anti-tnf-alpha bispecific antigen binding proteins and uses thereof

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims the benefit of Application No. PCT/US2016/052006, filed Sep. 15, 2016, and U.S. Provisional Application Nos.: 62/268,432, filed Dec. 16, 2015 and 62/333,063, filed May 6, 2016. Each of the foregoing applications is hereby incorporated by reference in its entirety. 
    
    
     FIELD OF THE INVENTION 
     This invention relates to biopharmaceuticals, particularly to therapeutic antigen binding proteins, methods of use thereof, pharmaceutical compositions thereof, and processes of making them. In particular, this invention relates to therapeutic antigen binding proteins that are capable of binding the cytokines TL1A and TNF-α. 
     BRIEF DESCRIPTION OF THE SEQUENCE LISTING 
     Incorporated herein by reference in its entirety is a Sequence Listing entitled, “A-1937-US-CNT_Seq_List_Corrections_012022_ST25_2136.txt”, comprising SEQ ID NO:1 through SEQ ID NO:2136, which includes nucleic acid and/or amino acid sequences disclosed herein. The Sequence Listing has been submitted herein in ASCII text format via EFS, and thus constitutes both the paper and computer readable form thereof. The Sequence Listing was first created using PatentIn on Nov. 22, 2016, and revised to remove errors on Jan. 20, 2021. This Sequence Listing is 3.543 MB in size. 
     BACKGROUND OF THE INVENTION 
     Cytokines are soluble, small proteins that mediate a variety of biological effects concerning the immune system. Such biological effects include induction of immune cell proliferation, development, differentiation, and/or migration; regulation of the growth and differentiation of many cell types; an inflammatory response through local or systemic accumulation of immune cells; and host-protective effects. See, for example, Arai et al.,  Annu. Rev. Biochem.  59:783 (1990); Mosmann,  Curr. Opin. Immunol.  3:311 (1991); Paul et al.,  Cell,  76:241 (1994)). Such immune effects can produce pathological consequences when the effect leads to excessive and/or chronic inflammation, as in autoimmune disorders (such as multiple sclerosis) and cancer/neoplastic diseases. Oppenheim et al., eds.,  Cytokine Reference,  Academic Press, San Diego, Calif. (2001); von Andrian et al.,  New Engl. J. Med.,  343:1020 (2000); Davidson et al.,  New Engl. J. Med.,  345:340 (2001); Lu et al.,  Mol. Cancer Res.,  4:221 (2006); Dalgleish et al.,  Cancer Treat Res.,  130:1 (2006). 
     TLIA (TNFSF15) is a cytokine, a TNF-α family member involved in T cell activation (Richard et al.,  J Leukoc Biol.  2015 September, 98(3):333-45). It is the ligand for Death Receptor 3 (DR3), also known as TNFRSF25. TL1A is mainly expressed at low basal level in monocytes, dendritic cells and endothelial cells, but highly induced after immune complex and cytokine and microbes stimulation. Multiple genome-wide association studies (GWAS) demonstrated TL1A single nucleotide polymorphisms (SNPs) associated with Crohn&#39;s disease in various ethnic populations. In addition, TL1A inflammatory bowel disease (IBD) risk SNPs were reported to be associated with Crohn&#39;s disease severity (Hirano,  IBD  19: 526, 2013). In the preclinical studies, TL1A protein treatment exacerbated colitis development in the colitis prone mdr1a−/− mice, but not in the wild-type mice. Altogether, human genomic data and preclinical colitis model data demonstrate that TL1A plays an important role in IBD development, and its blockage will be beneficial for IBD treatment. 
     Tumor Necrosis Factor-α (TNF-α) is a cytokine involved in the regulation of various physiological and pathological process (Buetler, et al.,  J Rheumatol Suppl.  57: 16-21, 1999). It is implicated in tumor regression, septic shock, cachexia, and a number of inflammatory and autoimmune conditions. Fransen et al. (June 1985), “Molecular cloning of mouse tumor necrosis factor cDNA and its eukaryotic expression,”  Nucleic Acids Res.  13 (12): 4417-29; Kriegler et al. (April 1988), “A novel form of TNF-α/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF-α,”  Cell  53 (1): 45-53. TNF-α inhibitors are a class of therapeutics approved to treat rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, plaque psoriasis, Crohn&#39;s disease, and ulcerative colitis (Sethi et al.,  Adv Exp Med Biol.  2009; 647:37-51). TNF-α inhibitors include etanercept, adalimumab, certolizumab pegol, infliximab, and golimumab. 
     Approximately 50% of patients respond to treatment with TNF inhibitors. However, only approximately 40% of those patients maintain responses after one year of treatment. Therefore, there is strong need for therapeutics with large effect size with durable response. We hypothesize that targeting multiple inflammatory pathways, such as TNF and TL1A, will likely achieve lamer effect size with anticipated safety profile. 
     SUMMARY OF THE INVENTION 
     This invention relates to the development of antigen binding proteins, particularly fully human antibodies that specifically bind TL1A. Preferred antigen binding proteins have strong binding affinity to both human and cynomolgus TL1A, and block TL1A mediated NF-kB activation and T cell activation. These antigen binding proteins have potential to be used as therapeutics for treatment of inflammatory bowel disease (IBD) and other autoimmune and inflammatory conditions. 
     The invention further relates to bispecific antigen binding proteins, particularly bispecific antibodies. Bispecific antigen binding proteins of the invention comprise a TL1A binding entity and a TNF-α binding entity. TL1A blocking proteins and TNF-α blocking proteins have different effects in studies measuring inhibition of induction of cytokines IFNγ, IL-5, IL-6, IL-8, and IL-10. TL1A but not TNF-α induces T cell activation in vivo. TL1A and TNF-α induce NF-κB in different cell types in human peripheral blood monocytes (PBMCs). TL1A and TNF-α further induce different cytokines in human PBMCs. TL1A and TNF-α induce some of the same genes but in different cell types (16 overlapping genes induced by TL1A in whole blood and TNF-α in NCM460 cells; 13 overlapping genes induced by TL1A in whole blood and TNF-α in PBMCs). There is a strong genetic association of TL1A with inflammatory bowel disease (IBD) and anti-TNF agents are clinically validated for treatment of IBD. Such bispecific antigen binding proteins thus are useful for treatment of IBD and other autoimmune and inflammatory conditions. For these and other reasons, there is a benefit to bispecific antigen binding proteins that specifically bind TL1A and TNF-α. 
     One format for such bispecific antigen binding proteins is heterodimeric immunoglobulins (hetero Ig). Such hetero Ig antigen binding proteins have one heavy chain-light chain pair directed to TL1A and another directed to TNF-α. 
     Another format for such bispecific antigen binding proteins is IgG-scFv molecules. In an IgG-scFv, each heavy chain of an antibody that specifically binds one target is linked to a single chain antibody (scFv) that specifically binds the other target. The scFv portion can be linked to the heavy chain of the IgG portion directly or through a peptide linker. In the IgG-scFv format, the IgG is directed to TL1A and the scFv portion to TNF-α or vice-versa. Bispecific antigen binding proteins in the IgG-scFv format have the advantage of being bivalent for both of their target antigens. 
     A further format for such bispecific antigen binding proteins is IgG-Fab molecules. In this format, the antigen binding protein has a structure such that a Fab molecule that binds to one target is fused to each heavy chain of an IgG molecule that binds to another target. One chain of a Fab portion can be linked directly or through a peptide linker to the C-terminus of a heavy chain of the IgG portion. The resulting molecule has the advantage of being tetravalent and bispecific. All variations of the IgG-Fab format preferably comprise the CDR sequences of Tables 21.2A and 21.2B hereinafter. Preferred heavy and light chain sequences of IgG-Fab molecules appear in Table 21.1 hereinafter. 
     Other formats for bispecific antigen binding proteins are within the scope of this invention. One such format is IgG-Fab molecules. In this format, each heavy chain of an antibody that specifically binds to one target is linked to a Fab fragment that specifically binds to the other target. In the IgG-Fab format, the IgG is directed to TL1A and the Fab portion to TNF-α or vice-versa. Bispecific antigen binding proteins in the IgG-Fab format have the advantage of being bivalent for both of their target antigens. Other formats for bispecific antigen binding proteins within the scope of this invention are described infra. 
     The invention also relates to isolated nucleic acids encoding the TL1A-specific antigen binding proteins of the invention, as well as vectors comprising the nucleic acids, host cells comprising the vectors, and methods of making and using the TL1A-specific antigen binding proteins. 
     The invention also relates to isolated nucleic acids encoding the bispecific antigen binding proteins of the invention, as well as vectors comprising the nucleic acids, host cells comprising the vectors, and methods of making and using the bispecific antigen binding proteins. 
     In other embodiments, the present invention provides compositions comprising the TL1A-specific antigen binding proteins, the bispecific antigen binding proteins and kits comprising the anti-TL1A or bispecific antigen binding proteins, as well as articles of manufacture comprising the anti-TL1A or bispecific antigen binding proteins. 
     The TL1A-specific antigen binding proteins and the bispecific antigen binding proteins described herein can be used in the manufacture of pharmaceutical compositions or medicaments for the treatment of conditions associated with TL1A and/or, in the case of bispecific antigen binding proteins, conditions associated with TNF-α. Thus, the present invention also provides pharmaceutical compositions comprising a TL1A-specific antigen binding protein or a bispecific antigen binding protein and a pharmaceutically acceptable diluent, excipient or carrier. 
     The invention further relates to methods of treatment using the TL1A-specific antigen binding proteins. The TL1A-specific antigen binding proteins of the present invention are useful for the inhibition of the proinflammatory cytokine TL1A. The antibodies can be used to reduce, limit, neutralize, or block the proinflammatory effects of TL1A. Thus, in some embodiments, the invention relates to the treatment of IBD and other autoimmune or inflammatory conditions using the TL1A-specific antigen binding proteins. 
     The invention further relates to methods of treatment using bispecific antigen binding proteins. The bispecific antigen binding proteins of the present invention are useful for the inhibition of the proinflammatory cytokines, TL1A and TNF-α. The bispecific binding proteins can be used to reduce, limit, neutralize, or block the proinflammatory effects of TL1A. Likewise, the bispecific antigen binding proteins hereof can be used to reduce, limit, neutralize, or block the proinflammatory effects of TNF-α. Thus, in some embodiments, the invention relates to the treatment of IBD and other autoimmune or inflammatory conditions using TL1A-specific/TNF-specific antigen binding proteins. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  shows a schematic representation of four bispecific hetero Ig formats used to generate anti-TL1A/anti-TNF-α bispecific antigen binding proteins. As shown, a heavy chain directed to one antigen is disulfide-bonded to a heavy chain directed to a different antigen. A light chain for each antigen is disulfide-bonded to the heavy chain for the corresponding antigen.  FIG. 1  shows a preferred embodiment, in which the heavy and light chains comprise charge mutations to aid in correct association of the heavy and light chains. The Kabat-Eu numbering scheme is used to denote the positions of charge pair mutations within each of the chains and for all sequences throughout this specification. This IgG-like bispecific antigen binding protein format is a heterotetramer comprising two different light chains and two different heavy chains. HC1 and LC1 refer to the heavy chain and light chain, respectively, of one Fab binding arm and HC2 and LC2 refers to the heavy chain and light chain, respectively, of the second Fab binding arm. For example, in the schematic, HC1 and LC1 correspond to the anti-TL1A receptor binding arm and HC2 and LC2 correspond to the anti-TNF-α binding arm. However, the two binding arms can be switched such that HC1 and LC1 correspond to the anti-TNF-α binding arm and HC2 and LC2 correspond to the anti-TL1A receptor binding arm. 
         FIG. 2  shows a schematic representation of the IgG-scFv format used to generate anti-TL1A/anti-TNF-α bispecific antigen binding proteins. As shown, the structure incorporates a full tetrameric IgG directed to one antigen. A single-chain variable fragment (scFv), which comprises variable domains from a second antibody linked together by a glycine-serine linker, is fused to the carboxyl terminus of the heavy chain of a first antibody through a peptide linker to produce a modified heavy chain. Although the VH-VL orientation of the variable domains within the scFv is shown, the variable domains may also be organized in a VL-VH orientation. The complete molecule is a multimer comprising two heavy chains (but one unique heavy chain sequence) and two light chains (but one unique light chain sequence) from the first antibody. 
         FIG. 3  concerns MabSelect SuRe affinity chromatography of an anti-TL1A/anti-TNF-α Hetero-Ig. It shows a representative FPLC protein A affinity capture chromatogram of an anti-TL1A/anti-TNF-α hetero-Ig. The protein was eluted with a step gradient of 100 mM acetic acid (conductivity: black trace, dashed), pH 3.6 and pooled based on the A280 (black trace, solid). 
         FIG. 4  shows a representative FPLC SP high performance sepharose purification chromatogram of an anti-TL1A/anti-TNF-α hetero-Ig. Protein was eluted with an increasing salt gradient (conductivity : black trace, dashed) and was pooled based on the A280 elution profile (black trace, solid) and Caliper LabChip analysis of fractions. 
         FIG. 5  shows a representative FPLC SP high performance sepharose purification chromatogram of an anti-TL1A/anti-TNF-α hetero-Ig. Protein was eluted with decreasing ammonium sulfate gradient (conductivity : black trace, dashed) and pooled based on the A280 elution profile (black trace, solid) and Caliper LabChip analysis of fractions. 
         FIG. 6  concerns Caliper analysis of a TL1A/TNF-α hetero-Ig. It shows non-reduced and reduced Caliper analysis of an anti-TL1A/anti-TNF-α hetero-Ig. 
         FIG. 7  concerns SE-HPLC analysis of an anti-TL1A/anti-TNF-α hetero-Ig. It shows size exclusion chromatography on 30 μg of the final anti-TL1A/anti-TNF-α hetero-Ig product injected onto a Sepax Zenix-C SEC-300 column (7.8×300 mm) in 50 mM NaH 2 PO 4 , 250 mM NaCl, pH 6.9 at 1 ml/min, observing the absorbance at 280 nm (black trace). 
         FIG. 8  concerns LC-MS of Non-Reduced Hetero-Ig (Theoretical mass: 145495 Da).  FIG. 8  shows mass analysis of 20 pg non-reduced anti-TL1A/anti-TNF-α hetero-Ig eluted from a reverse-phase HPLC gradient using an Agilent Zorbax 300SB-C8 column (2.1×50 mm 3.5 μm) and mobile phases of 0.1% TFA and 90% n-Propanol/0.1% TFA (mobile phases A and B, respectively), equipped with an Agilent 6230 ESI-TOF Mass Spectrometer. 
         FIG. 9  shows mass analysis of 20 μg TL1A/TNF-α hetero-Ig after limited Lysyl Endoproteinase C digestion for 30 minutes in 100 mM TRIS, pH 8. Reverse-phase HPLC was conducted using an Agilent Zorbax 300SB-C8 column (2.1×50 mm 3.5 μm) and mobile phases of 0.1% TFA and 90% n-propanol/0.1% TFA (mobile phases A and B respectively) and mass detection on an Agilent 6230 ESI-TOF Mass Spectrometer. Theoretical masses for TL1A Fab, TNF Fab, and Fc are 47350 Da, 48002 Da, and 50175 Da, respectively. 
         FIG. 10  concerns MabSelect SuRe affinity chromatography of a anti-TL1A/anti-TNF-α IgG-scFv. It shows a representative FPLC protein A affinity capture chromatogram of an anti-TL1A/anti-TNF-α IgG-scFv. The protein was eluted with a step gradient of 100 mM acetic acid, pH 3.6 and pooled based on the A280 (black trace). 
         FIG. 11  concerns SP Sepharose High Performance chromatography of an anti-TL1A/anti-TNF-α IgG-scFv. It shows a representative FPLC Superdex 200 purification chromatogram of an anti-TL1A/anti-TNF-α IgG-scFv. Protein was eluted with isocratic gradient of buffer and pooled based on the A280 elution profile (black trace) and SE-HPLC analysis of fractions. 
         FIG. 12  shows non-reduced and reduced Caliper analysis of an anti-TL1A/anti-TNF-α IgG-scFv. 
         FIG. 13  concerns SE-HPLC analysis of an anti-TL1A/anti-TNF-α IgG-scFv. It shows size exclusion chromatography on 30 μg of the final anti-TL1A/anti-TNF-α IgG-scFv product injected onto a Sepax Zenix-C SEC-300 column (7.8×300 mm) in 50 mM NaH 2 PO 4 , 250 mM NaCl, pH 6.9 at 1 ml/min, observing the absorbance at 280 nm. 
         FIG. 14  concerns IdeS Protease digested Ig-scFv. It shows mass analysis of 20 μg Ig-scFv using reverse-phase HPLC separation on an Agilent Zorbax 300SB column (2.1×50 mm, 3.5 μm) with mobile phases of 0.1% TFA and 90% n-propanol/0.1% TFA (mobile phases A and B, respectively), and detection on an Agilent 6230 ESI-TOF Mass Spectrometer. 
         FIG. 15  shows genotyping of TL1A SNPs. To evaluate potential TL1A genotype association with expression, genomic DNA (gDNA) was isolated from healthy PBMC donors using Gentra Puregene Tissue kit from Qiagen. Genomic DNA were genotyped using TaqMan SNP genotyping assays for rs7848647, rs6478109, rs6478108, and rs3810936 assays from LifeTech and standard protocols on the Bio-Rad droplet digital PCR platform. Donors were considered homozygous risk haplotype if only risk alleles were present at all 4 genotyped SNPs (rs7848647, rs6478109, rs6478108, and rs3810936). Donors were considered homozygous non-risk haplotype if only non-risk alleles were present at all 4 genotyped SNPs. Donors were considered heterozygous haplotype if both risk and non-risk alleles were present at all 4 genotyped SNPs. Donors that were considered “recombinant” had only homozygous risk alleles at rs7848647, rs6478109, and rs6478108, but were heterozygous (risk and non-risk alleles present) at rs3810936. 
         FIGS. 16A and 16B  show higher fold induction of TL1A risk allele than non-risk allele in a heterozygous PBMC AEI Study. Frequency of risk vs. non-risk allele usage in heterozygous PBMC at basal level or after immune complex stimulation at various time points was examined by droplet digit PCR (ddPCR) using synonymous SNP (rs3810936) allelic specific fluorescent probes. The allelic expression ratio was calculated by dividing the copies/ml of the risk allele by the copies/ml of the non-risk allele. Total copy number of each allele was normalized by the input amount of cDNA (copies/ng), and the fold-induction for each allele at each time point was calculated by dividing the copies/ng at the time point of interest by the copies/ng at baseline (0 hr). 
         FIG. 17  shows that TL1A SNPs in promoter and intron contribute to allelic expression imbalance regulation. The inventor(s) identified a small number of donors homozygous for risk alleles at rs7848647, rs6478109, and rs6478108, but heterozygous at rs3810936, likely due to recombination between rs6478109 and synonymous SNP rs3810936. Frequency of risk vs. non-risk allele usage in heterozygous or recombinant donor PBMC was examined by droplet digit PCR (ddPCR) using synonymous SNP (rs3810936) allelic specific fluorescent probes. Compared to heterozygous donors, no allelic expression imbalance was detected in these recombinant individuals before or after immune complex stimulation. 
         FIGS. 18A and 18B  show that TL1A risk SNPs are associated with expression quantitative trait loci (eQTL), PBMC donors with homozygous TL1A risk allele, homozygous non-risk alleles or heterozygous alleles were treated with immune complex. Total expression of TL1A (TNFSF15) in each donor was determined by digital PCR. In brief, cDNA from each sample was mixed with ddPCRTM Supermix for Probes (Bio-Rad) and PrimeTime® qPCR assay ID Hs.PT.56a.41003970. Droplets were generated for each reaction using the QX100™ droplet generator (Bio-Rad) and subjected to thermal cycling on a C1000 Touch™ thermal cycler (Bio-Rad). Following amplification, droplet fluorescence was read on a QX100™ droplet reader (Bio-Rad). Data were analyzed using QuantaSoft software (Bio-Rad) and copies/μl of TL1A was determined and normalized for the amount of input cDNA (copies/ng). P values for differences in TL1A expression levels between different TNFSF15 haplotypes were determined using the student t test. At basal level, PBMCs from donors homozygous for TL1A risk SNPs have lower TL1A mRNA compared to non-risk homozygous donors. However, after immune complex stimulation, PBMCs from donors homozygous for TL1A risk SNPs have higher TL1A expression compared to non-risk homozygous donors. 
         FIGS. 19A and 19B  show cell-type specific regulation of TL1A synonymous SNP allelic expression imbalance. HUVEC cells from various donors were genotyped as described previously. Genotyped HUVEC cells from various donors were treated with IL-1 at various time points. Total copy number of each allele was measured as described previously. The allelic expression ratio was calculated by dividing the copies/ml of the risk allele by the copies/ml of the non-risk allele. No allelic expression imbalance was observed in HUVEC cells with or without IL-1 treatment. 
         FIGS. 20A and 20B  show that prophylactivc treatment with anti-TL1A monoclonal antibody inhibited spontaneous colitis development in mdr1a −/−  mice. Mice (n=10, 6-7 weeks age) were randomized to different groups and treated intra-peritoneally once a week with 500 μg of anti-mouse TL1A antibody, anti-IL23p19 antibody, anti-mouse IL17RA antibody, or mouse isotype control or no treatment once per week for 8 weeks. Clinical disease activity was monitored by evaluating anal inflammation and stool consistency. Sections of the intestine were subjected to histopath analysis. Statistical analysis was performed using one-way ANOVA with Dunett&#39;s compared to mIgG1 group. 
         FIGS. 21A and 21B  show that TL1A exacerbated colitis in mdr1a +  mice. Mdr1a −/−  mice or wild-type control mice at 6-8 weeks of age were treated intra-peritoneally once a week with 150 μg of recombinant mFc-TL1A fusion protein or isotype control three times each week for 4 weeks. Clinical disease activity was monitored by evaluating anal inflammation and stool consistency as described previously. After 4 weeks of treatment, mice were sacrificed, sections of the intestine were were subjected to histopath analysis. Statistical analysis was performed using one way ANOVA with Dunett&#39;s compared to mIgG1 group. 
         FIGS. 22A to 22F  shows increased inflammatory cells in lamina propria from Fc-TL1A challenged mdr1a −/−  mice. Mdr1a −/−  mice or wild-type control mice at 6-8 weeks of age were treated intra-peritoneally once a week with 150 μg of recombinant mFc-TL1A fusion protein or isotype control three times each week for 4 weeks. Lamina propria lymphocytes were isolated and stained for surface antigens and analyzed by FACS. TL1A challenge in mdr1a −/−  mice resulted in increased inflammatory cells in lamina propria. 
         FIGS. 23A to 23E  show distinct cytokine induction by TL1A and TNF challenge in mice. To evaluate if TL1A and TNF challenge result in similar or different pharmacodynamics effects, C57Bl/6 mice (8 week, female) were first intraperitonially injected with 500 μg/mice of anti mouse TL1A, anti mouse TNF-α or PBS. After 4 hours, the mice were then challenged with 100 μg/mice of TL1A or 10 μg/mice of TNF-α or without challenging. The sera were collected after 24 hours. The cytokines were measured by MSD (IL-22 was measured by ELISA). 
         FIGS. 24A to 24F  shows distinct cytokine induction by TL1A and TNF treatment in human PBMC. PBMC freshly isolated from human blood were cultured in media (RPMI1640 supplemented with 10% FBS, 2 mM glutamine, 1 mM sodium pyruvate, 5×10 −5  M 2-ME, and antibiotics) in the presence of 100 ng/ml of human TL1A or TNF-α. Supernatant was collected after 72 hours. The cytokines in the supernatant were measured by MSD (IL-22 was measured by ELISA). 
         FIG. 25  depicts a schematic representation of a bispecific IgG-Fab format for an anti-TL1A anti-TNF-α bispecific antigen binding protein within the present invention. In this format, one polypeptide chain of a Fab fragment from a second antibody (e.g. the heavy chain (VH2-CH1) is fused to the carboxyl terminus of each heavy chain of a first antibody through a peptide linker to produce a modified heavy chain. The complete molecule is a homohexamer comprising two modified heavy chains, two light chains from the first antibody, and two polypeptide chains containing the other half of the Fab fragment from the second antibody (e.g. the light chain (VL2-CL)). Charge pair mutations (represented by the circles) can be introduced into the Fab regions of the first antibody (Fab 1) and/or second antibody (Fab 2) to promote correct heavy chain-light chain pairing. 
         FIG. 26  depicts a schematic representation of a bispecific IgG-Fab format for an anti-TL1A anti-TNF-α bispecific antigen binding protein within the present invention using immunoglobulin domain crossover. In this variation of the IgG-Fab format, one polypeptide chain of a Fab fragment from a second antibody (e.g. the heavy chain (VH2-CH1) is fused to the carboxyl terminus of the heavy chain comprising a CL instead of a CH1 domain of a first antibody through a peptide linker to produce a modified heavy chain. In this way, the CH1 and the CL domains of Fab 1 are “swapped.” This swap is referred to as a Fab1 swap or an N-terminal swap herein. The complete molecule is a homohexamer comprising two modified heavy chains, two light chains from the first antibody comprising a CH1 domain instead of a CL domain, and two polypeptide chains containing the other half of the Fab fragment from the second antibody (e.g. the light chain (VL2-CL)). Charge pair mutations (represented by the circles) can be introduced into the Fab regions of the first antibody (Fab 1) and/or second antibody (Fab 2) to promote correct heavy chain-light chain pairs. Not shown but also within the scope of this invention are IgG-Fab molecules in which (i) the CH1 and CL domains of Fab 2 are swapped instead of those of Fab 1 (referred to herein as a Fab 2 swap or a C-terminal swap) or (ii) both the CH1 and CL domains of Fab 1 are swapped and the CH1 and CL domains of Fab 2 are swapped (referred to herein as a dual swap). 
         FIG. 27  compares the expression titer of IgG-Fabs based on domain swapping format. 
         FIG. 28  compares the expression titer of IgG-Fabs based on type of charge pair mutation(s). 
         FIG. 29  compares the purity of IgG-Fabs based on domain swapping format. 
         FIG. 30  compares the anti-TL1A potency of IgG-Fabs based on domain swapping format. 
         FIG. 31  compares the anti-TNF-α potency of IgG-Fabs based on domain swapping format. 
         FIG. 32  compares the anti-TL1A potency of IgG-Fabs based on type of charge pair mutation(s). 
         FIG. 33  compares the anti-TNF-α potency of IgG-Fabs based on type of charge pair mutation(s). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     Definition of Terms 
     In the description that follows, a number of terms are used extensively. The following definitions are provided to facilitate understanding of the invention. 
     Unless otherwise specified, “a”, “an”, “the”, and “at least one” are used interchangeably and mean one or more than one. 
     A “binding entity” as used herein means any monomeric or multimeric protein or protein fragment that specifically binds a specified target antigen. The term “binding entity” includes but is not limited to antibodies and binding parts thereof, such as immunologically functional fragments. Peptibodies and peptides are other examples of binding entities. The term “immunologically functional fragment” (or simply “fragment”) of an antibody or immunoglobulin chain (heavy or light chain) binding entity, as used herein, is a species of binding entity comprising a portion (regardless of how that portion is obtained or synthesized) of an antibody that lacks at least some of the amino acids present in a full-length chain but which is still capable of specifically binding to an antigen. Such fragments are biologically active in that they bind to the target antigen and can compete with other binding entities, including intact antibodies, for binding to a given epitope. In some embodiments, the fragments are neutralizing fragments. In some embodiments, the fragments can block or reduce the likelihood of the interaction between the target antigen (TL1A or TNF-α) and its receptor. In one aspect, such a fragment will retain at least one CDR present in the full-length light or heavy chain, and in some embodiments will comprise a single heavy chain and/or light chain or portion thereof. These biologically active fragments can be produced by recombinant DNA techniques, or can be produced by enzymatic or chemical cleavage of binding entities, including intact antibodies. Immunologically functional immunoglobulin fragments include, but are not limited to, Fab, a diabody (heavy chain variable domain on the same polypeptide as a light chain variable domain, connected via a short peptide linker that is too short to permit pairing between the two domains on the same chain), Fab′, F(ab′) 2 , Fv, domain antibodies and single-chain antibodies, and can be derived from any mammalian source, including but not limited to human, mouse, rat, camelid or rabbit. It is further contemplated that a functional portion of the binding entities disclosed herein, for example, one or more CDRs, could be covalently bound to a second protein or to a small molecule to create a therapeutic agent directed to a particular target in the body, possessing bifunctional therapeutic properties, or having a prolonged serum half-life. As will be appreciated by one of skill in the art, a binding entity can include non-protein components. In some sections of the present disclosure, examples of binding entities are named herein in terms of “number/letter/number” (e.g., 23B3), with some binding entities further identified by additional letter/number combinations (e.g., VH4). In these cases, the exact name denotes a specific antibody. That is, a binding entity named 23B3 may have some degree of sequence identity with but is not the same as an antibody named 23B3 VH4 (unless they are explicitly taught to be the same in the specification). 
     A “TL1A binding entity” is a binding entity that specifically binds to human TL1A; i.e., a binding entity for which human TL1A is the target antigen. 
     A “TNF-α binding entity” is a binding entity that specifically binds to human TNF-α; i.e., a binding entity for which human TNF-α is the target antigen. 
     “Antigen binding protein” refers to a protein or polypeptide that comprises an antigen-binding region or antigen-binding portion that has a strong affinity for another molecule to which it binds (antigen). Antigen-binding proteins encompass antibodies, peptibodies, antibody fragments, antibody derivatives, antibody analogs, fusion proteins, and antigen receptors including chimeric antigen receptors (CARs). 
     “Antibodies” (Abs) and “immunoglobulins” (Igs) are glycoproteins having the same structural characteristics. While antibodies exhibit binding specificity to a specific antigen, immunoglobulins include both antibodies and other antibody-like molecules that lack antigen specificity. Polypeptides of the latter kind are, for example, produced at low levels by the lymph system and at increased levels by myelomas. Thus, as used herein, the term “antibody” or “antibody peptide(s)” refers to an intact antibody, an antibody that competes for specific binding with an antibody disclosed in this specification, or an antigen-binding fragment thereof that competes with the intact antibody for specific binding and includes chimeric, humanized, fully human, and bispecific antibodies. In certain embodiments, antigen-binding fragments are produced, for example, by recombinant DNA techniques. In additional embodiments, antigen-binding fragments are produced by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include, but are not limited to, Fab, Fab′, F(ab) 2 , F(ab′) 2 , Fv, and single-chain antibodies. 
     The term “isolated antibody” as used herein refers to an antibody that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials which would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the antibody will be purified (1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody&#39;s natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step. 
     The term “agonist” refers to any compound including a protein, polypeptide, peptide, antibody, antibody fragment, large molecule, or small molecule (less than 10 kD), that increases the activity, activation or function of another molecule. 
     The term “antagonist” refers to any compound including a protein, polypeptide, peptide, antibody, antibody fragment, large molecule, or small molecule (less than 10 kD), that decreases the activity, activation or function of another molecule. 
     The term “bind(ing)” of an antigen or other polypeptide includes, but is not limited to, the binding of a ligand polypeptide of the present invention to a receptor; the binding of a receptor polypeptide of the present invention to a ligand; the binding of an antibody of the present invention to an antigen or epitope; the binding of an antigen or epitope of the present invention to an antibody; the binding of an antibody of the present invention to an anti-idiotypic antibody; the binding of an anti-idiotypic antibody of the present invention to a ligand; the binding of an anti-idiotypic antibody of the present invention to a receptor; the binding of an anti-anti-idiotypic antibody of the present invention to a ligand, receptor or antibody, etc. 
     A “bispecific” antigen binding protein is a molecule with binding entities derived from both a first antigen binding protein (e.g., antibody) that specifically binds a first target molecule of interest and a second antigen binding protein (e.g., antibody) that specifically binds a second target molecule of interest. Bispecific antigen binding proteins may be produced by a variety of methods including, but not limited to, fusion of hybridomas or linking of Fab′ fragments. See, e.g., Songsivilai et al., Clin. Exp. Immunol., 79:315-321 (1990); Kostelny et al., J. Immunol., 148:1547-1553 (1992), hereby incorporated by reference. Molecules in the formats depicted in  FIGS. 1 and 2  are bispecific antigen binding proteins in accordance with this definition. Various additional formats of bispecific antigen binding proteins within this definition are disclosed in WO 2014/159725; WO 2013/041687; U.S. Pat. Nos. 8,945,553; 8,945,553; 8,258,268; US Pat. App. No. 2012/0195900; International patent application WO 2012/088302; U.S. Prov. App. 62/218,977; Fischer and Leger (2007),  Pathobiology  74:3-14; van Spriel et al. (2000),  Immunology Today  21(8): 391-7; Kufer et al. (2004),  Trends in Biotechnology  22(5): 238-44; Byrne et al. (2013),  Trends in Biotechnology  31(11): 621-31; Muller and Kontermann (2010),  Biodrugs  24(2): 89-98; Chames and Baty (2009), http://dx.doi.org/10.4161/mabs.1.6.10015; Kontermann (2012), http://dx.doi.org/10.4161/mabs.4.2.19000; Holliger et al. (1993),  Proc. Natl. Acad. Sci. USA  90: 6444-8; Speiss et al.,  Mol. Immunol.  (2015), 67: 95-106, http://dx.doi.org/10.1016/j.molimm.2015.01.003; Holliger et al. (1993),  Proc. Natl. Acad. Sci. USA  90: 6444-8; Speiss et al.,  Mol. Immunol.  (2015), http://dx.doi.org/10.1016/j.molimm.2015.01.003; Ridgway et al, (1996),  Protein Eng.  9: 617; Gunasekaran et al (2010),  J. Biol. Chem.  285:19637; Davis (2010),  Protein Eng. Des.  &amp;  Sel.  23:195; DiGiammarino et al. (2012),  Methods Mol. Biol.  899:145, 2012); Lindhofer et al. (1995),  J. Immunol.  155: 219: Schaefer et al. (2011),  Proc. Natl. Acad. Sci. USA,  108: 11187; Regula et al, US Patent Application No: 2010/0322934; Bostrom et al. (2009),  Science  323:1610; US Pat. App. 2011/0076722; Rossi et al. (2008),  Cancer Res.  68:8384; each of which is hereby incorporated by reference. 
     The term “epitope” refers to the portion of an antigen to which an antibody specifically binds. Thus, the term “epitope” includes any protein determinant capable of specific binding to an immunoglobulin or T-cell receptor. Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. More specifically, the term “IL-17 epitope”, “TNF-α epitope” and/or “TNF-α/p19 epitope” as used herein refers to a portion of the corresponding polypeptide having antigenic or immunogenic activity in an animal, preferably in a mammal, and most preferably in a mouse or a human. An epitope having immunogenic activity is a portion of, for example, an IL-17A or IL-17F or TNF-α/p19 polypeptide that elicits an antibody response in an animal. An epitope having antigenic activity is a portion of, for example, an IL-17A or IL-17F or TNF-α/p19 polypeptide to which an antibody immunospecifically binds as determined by any method well known in the art, for example, by immunoassays, protease digest, crystallography or HID-Exchange. Antigenic epitopes need not necessarily be immunogenic. Such epitopes can be linear in nature or can be a discontinuous epitope. Thus, as used herein, the term “conformational epitope” refers to a discontinuous epitope formed by a spatial relationship between amino acids of an antigen other than an unbroken series of amino acids. 
     The term “immunoglobulin” refers to a protein consisting of one or more polypeptides substantially encoded by immunoglobulin genes. One form of immunoglobulin constitutes the basic structural unit of an antibody. This form is a tetramer and consists of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions are together responsible for binding to an antigen, and the constant regions are responsible for the antibody effector functions. 
     Full-length immunoglobulin “light chains” (about 25 Kd or about 214 amino acids) are encoded by a variable region gene at the NH2-terminus (about 110 amino acids) and a kappa or lambda constant region gene at the COOH-terminus. Full-length immunoglobulin “heavy chains” (about 50 Kd or about 446 amino acids), are similarly encoded by a variable region gene (about 116 amino acids) and one of the other aforementioned constant region genes (about 330 amino acids). Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, and define the antibody&#39;s isotype as IgG (such as IgG1, IgG2, IgG3 and IgG4), IgM, IgA, IgD and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. (See generally, Fundamental Immunology (Paul, W., ed., 2nd Edition, Raven Press, NY (1989)), Chapter 7 (incorporated by reference in its entirety for all purposes). 
     An immunoglobulin light or heavy chain variable region consists of a “framework” region interrupted by three hypervariable regions. Thus, the term “hypervariable region” refers to the amino acid residues of an antibody which are responsible for antigen binding. The hypervariable region comprises amino acid residues from a “Complementarity Determining Region” or “CDR” (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and/or those residues from a “hypervariable loop” (Chothia et al., J. Mol. Biol. 196: 901-917 (1987)) (both of which are incorporated herein by reference). “Framework Region” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined. The sequences of the framework regions of different light or heavy chains are relatively conserved within a species. Thus, a “human framework region” is a framework region that is substantially identical (about 85% or more, usually 90-95% or more) to the framework region of a naturally occurring human immunoglobulin. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDR&#39;s. The CDR&#39;s are primarily responsible for binding to an epitope of an antigen. Accordingly, the term “humanized” immunoglobulin refers to an immunoglobulin comprising a human framework region and one or more CDR&#39;s from a non-human (usually a mouse or rat) immunoglobulin. The non-human immunoglobulin providing the CDR&#39;s is called the “donor” and the human immunoglobulin providing the framework is called the “acceptor”. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, preferably about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDR&#39;s, are substantially identical to corresponding parts of natural human immunoglobulin sequences. Further, one or more residues in the human framework region may be back mutated to the parental sequence to retain optimal antigen-binding affinity and specificity. In this way, certain framework residues from the non-human parent antibody are retained in the humanized antibody in order to retain the binding properties of the parent antibody while minimizing its immunogenicity. The term “human framework region” as used herein includes regions with such back mutations. A “humanized antibody” is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin. For example, a humanized antibody would not encompass a typical chimeric antibody as defined above, e.g., because the entire variable region of a chimeric antibody is non-human. 
     As used herein, the term “modified heavy chain” refers to a fusion protein comprising an immunoglobulin heavy chain, particularly a human IgG1 or human IgG2 heavy chain, and a functional antibody fragment (e.g., a Fab or scFv) or portion thereof (e.g. immunoglobulin light chain or Fd fragment), wherein the fragment or portion thereof is fused at its N-terminus, optionally through a peptide linker, to the C-terminus of the heavy chain. 
     The term “humanized” immunoglobulin refers to an immunoglobulin comprising a human framework region and one or more CDR&#39;s from a non-human, e.g., mouse, rat or rabbit, immunoglobulin. The non-human immunoglobulin providing the CDR&#39;s is called the “donor” and the human immunoglobulin providing the framework is called the “acceptor”. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, preferably about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDR&#39;s and possibly a few back-mutated amino acid residues in the framework region (e.g., 1-15 residues), are substantially identical to corresponding parts of natural human immunoglobulin sequences. A “humanized antibody” is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin. For example, a humanized antibody would not encompass a typical chimeric antibody as defined above, e.g., because the entire variable region of a chimeric antibody is non-human. 
     The terms “human antibody” and “fully human antibody” each refer to an antibody that has an amino acid sequence of a human immunoglobulin, including antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins and that do not express endogenous immunoglobulins; for example, Xenomouse antibodies and antibodies as described by Kucherlapati et al. in U.S. Pat. No. 5,939,598. 
     The term “genetically altered antibodies” means antibodies wherein the amino acid sequence has been varied from that of a native antibody. Because of the relevance of recombinant DNA techniques in the generation of antibodies, one need not be confined to the sequences of amino acids found in natural antibodies; antibodies can be redesigned to obtain desired characteristics. The possible variations are many and range from changing just one or a few amino acids to complete redesign of, for example, the variable and/or constant region. Changes in the constant region will, in general, be made in order to improve or alter characteristics, such as complement fixation, interaction with membranes and other effector functions. Changes in the variable region will be made in order to improve the antigen binding characteristics. 
     A “Fab fragment” is comprised of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule. 
     A “Fab′ fragment” contains one light chain and one heavy chain that contains more of the constant region, between the C H1  and C H2  domains, such that an interchain disulfide bond can be formed between two heavy chains to form a F(ab′) 2  molecule. 
     A “F(ab′) 2  fragment” contains two light chains and two heavy chains containing a portion of the constant region between the C H1  and C H2  domains, such that an interchain disulfide bond is formed between two heavy chains. 
     The term “native Fc” refers to molecule or sequence comprising the sequence of a non-antigen-binding fragment resulting from digestion of whole antibody, whether in monomeric or multimeric form. The original immunoglobulin source of the native Fc is preferably of human origin and may be any of the immunoglobulins, although IgG1, IgG2 and IgG4 are preferred. Native Fc&#39;s are made up of monomeric polypeptides that may be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and non-covalent association. The number of intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from 1 to 4 depending on class (e.g., IgG, IgA, IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, IgGA2). One example of a native Fc is a disulfide-bonded dimer resulting from papain digestion of an IgG (see Ellison et al. (1982), Nucleic Acids Res. 10: 4071-9). The term “native Fc” as used herein is generic to the monomeric, dimeric, and multimeric forms. 
     The term “Fc variant” refers to a molecule or sequence that is modified from a native Fc but still comprises a binding site for the salvage receptor, FcRn. International applications WO 97/34631 (published 25 Sep. 1997) and WO 96/32478 describe exemplary Fc variants, as well as interaction with the salvage receptor, and are hereby incorporated by reference. Thus, the term “Fc variant” comprises a molecule or sequence that is humanized from a non-human native Fc. Furthermore, a native Fc comprises sites that may be removed because they provide structural features or biological activity that are not required for the fusion molecules of the present invention. Thus, the term “Fc variant” comprises a molecule or sequence that lacks one or more native Fc sites or residues that affect or are involved in (1) disulfide bond formation, (2) incompatibility with a selected host cell (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC). Fc variants are described in further detail hereinafter. 
     The term “Fc domain” encompasses native Fc and Fc variant molecules and sequences as defined above. As with Fc variants and native Fc&#39;s, the term “Fc domain” includes molecules in monomeric or multimeric form, whether digested from whole antibody or produced by other means. 
     The term “multimer” as applied to Fc domains or molecules comprising Fc domains refers to molecules having two or more polypeptide chains associated covalently, noncovalently, or by both covalent and non-covalent interactions. IgG molecules typically form dimers; IgM, pentamers; IgD, dimers; and IgA, monomers, dimers, trimers, or tetramers. Multimers may be formed by exploiting the sequence and resulting activity of the native Ig source of the Fc or by derivatizing (as defined below) such a native Fc. 
     The term “dimer” as applied to Fc domains or molecules comprising Fc domains refers to molecules having two polypeptide chains associated covalently or non-covalently. Thus, exemplary dimers within the scope of this invention are as shown in  FIG. 2 . 
     The terms “Fv fragment” and “single chain antibody” refer to polypeptides containing antibody variable regions from both heavy and light chains but lacking constant regions. Like a whole antibody, it is able to bind selectively to a specific antigen. With a molecular weight of only about 25 kDa, Fv fragments are much smaller than common antibodies (150-160 kD) which are composed of two heavy protein chains and two light chains, and even smaller than Fab fragments (about 50 kDa, one light chain and half a heavy chain). 
     A “single domain antibody” is an antibody fragment consisting of a single domain Fv unit, e.g., V H  or V L . Like a whole antibody, it is able to bind selectively to a specific antigen. With a molecular weight of only 12-15 kDa, single-domain antibodies are much smaller than common antibodies (150-160 kDa) which are composed of two heavy protein chains and two light chains, and even smaller than Fab fragments (.about.50 kDa, one light chain and half a heavy chain) and single-chain variable fragments (.about.25 kDa, two variable domains, one from a light and one from a heavy chain). The first single-domain antibodies were engineered from heavy-chain antibodies found in camelids. Although most research into single-domain antibodies is currently based on heavy chain variable domains, light chain variable domains and nobodies derived from light chains have also been shown to bind specifically to target epitopes. 
     The term “monoclonal antibody” as used herein is not limited to antibodies produced through hybridoma technology. The term “monoclonal antibody” refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced. 
     In some embodiments, the anti-TL1A antigen binding protein, bispecific antigen binding protein or functional fragment of either thereof from which the anti-TL1A binding domain is derived selectively inhibits human TL1A relative to TNF superfamily ligands. An antibody or functional fragment thereof “selectively inhibits” a specific receptor or ligand relative to other receptors or ligands when the IC50 of the antibody in an inhibition assay of the specific receptor is at least 50-fold lower than the IC50 in an inhibition assay of another “reference” ligand or receptor. An “IC50” is the dose/concentration required to achieve 50% inhibition of a biological or biochemical function. With radioactive ligands, IC50 is the concentration of a competing ligand that displaces 50% of the specific binding of the radioactive ligand. The IC50 of any particular substance or antagonist can be determined by constructing a dose-response curve and examining the effect of different concentrations of the drug or antagonist on reversing agonist activity in a particular functional assay. IC50 values can be calculated for a given antagonist or drug by determining the concentration needed to inhibit half of the maximum biological response of the agonist. Thus, the IC50 value for any anti-TL1A antibody or functional fragment thereof can be calculated by determining the concentration of the antibody or fragment needed to inhibit half of the maximum biological response of TL1A in activating the human TL1A receptor in any functional assay, such as the assay described in Example 14 hereinafter. An antibody or functional fragment thereof that selectively inhibits a specific ligand or receptor is understood to be a neutralizing antibody or neutralizing fragment with respect to that ligand or receptor. Thus, in some embodiments, the anti-TL1A antibody or functional fragment thereof from which the anti-TL1A binding domain of the bispecific antigen binding proteins of the invention is derived is a neutralizing antibody or fragment of human TL1A. 
     The variable regions or CDR regions of any anti-TL1A antibody or functional fragment thereof can be used to construct the anti-TL1A binding entity of any of the bispecific antigen binding proteins described herein. Likewise, the variable regions or CDR sequences of any anti-TNF-α antibody or functional fragment thereof can be used to construct the anti-TNF-α binding entity of any of the bispecific antigen binding proteins described herein. For instance, the anti-TL1A binding domain of the bispecific antigen binding proteins of the invention may comprise VH and/or VL regions or one or more CDRs from any of the anti-human TL1A antibodies described in U.S. Pat. No. 7,820,798; US Pat. App. 2008/0003221; U.S. Pat. No. 8,263,743; US Pat. App. 2011/0217310; US Pat. App. 2012/0263718; US Pat. App. 2014/0308271; WO 2012/161856; WO 2013/044298; WO 2005/018571; US Pat. App. 2014/0120109; U.S. Pat. No. 6,521,422; US Pat. App. 2014/0255302; and US Pat. App. 2015/0132311; each of which is hereby incorporated by reference in its entirety. In some embodiments, the anti-TL1A antibody from which the anti-TL1A binding entity is derived cross-blocks one or more of the human anti-TL1A antibodies described in one of the references just mentioned or one or more of the human anti-TL1A antibodies described below. The terms “cross-block,” “cross-blocked,” and “cross-blocking” are used interchangeably herein to mean the ability of an antibody to interfere with the binding of other antibodies or binding fragments to a target (e.g. human TL1A). The extent to which an antibody or binding fragment is able to interfere with the binding of another to a target (e.g., human TL1A) and therefore whether it can be said to cross-block, can be determined using competition binding assays. In some embodiments, a cross-blocking antibody or binding fragment thereof reduces human TL1A binding of a reference antibody between about 40% and 100%, such as about 60% and about 100%, specifically preferably between about 70% and 100%, and more specifically preferably between about 80% and 100%. A particularly suitable quantitative assay for detecting cross-blocking uses a Biacore machine which measures the extent of interactions using surface plasmon resonance technology. Another suitable quantitative cross-blocking assay uses a FACS-based approach to measure competition between antibodies in terms of their binding to human TL1A. An exemplary assay regarding such cross-blocking appears in US Pat. App. 2015/0132311, example 2, paragraphs [0922]-[0924], hereby incorporated by reference. 
     The term “nucleic acid” or “nucleic acid molecule” refers to polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action. Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., .alpha.-enantiomeric forms of naturally-occurring nucleotides), or a combination of both. Modified nucleotides can have alterations in sugar moieties and/or in pyrimidine or purine base moieties. Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters. Moreover, the entire sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs. Examples of modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes. Nucleic acid monomers can be linked by phosphodiester bonds or analogs of such linkages. Analogs of phosphodiester linkages include phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate, phosphoramidate, and the like. The term “nucleic acid molecule” also includes so-called “peptide nucleic acids”, which comprise naturally-occurring or modified nucleic acid bases attached to a polyamide backbone. Nucleic acids can be either single stranded or double stranded. 
     The term “complement of a nucleic acid molecule” refers to a nucleic acid molecule having a complementary nucleotide sequence and reverse orientation as compared to a reference nucleotide sequence. 
     The term “degenerate nucleotide sequence” denotes a sequence of nucleotides that includes one or more degenerate codons as compared to a reference nucleic acid molecule that encodes a polypeptide. Degenerate codons contain different triplets of nucleotides, but encode the same amino acid residue (i.e., GAU and GAC triplets each encode Asp). 
     An “isolated nucleic acid molecule” is a nucleic acid molecule that is not integrated in the genomic DNA of an organism. For example, a DNA molecule that encodes a growth factor that has been separated from the genomic DNA of a cell is an isolated DNA molecule. Another example of an isolated nucleic acid molecule is a chemically-synthesized nucleic acid molecule that is not integrated in the genome of an organism. A nucleic acid molecule that has been isolated from a particular species is smaller than the complete DNA molecule of a chromosome from that species 
     A “nucleic acid molecule construct” is a nucleic acid molecule, either single- or double-stranded, that has been modified through human intervention to contain segments of nucleic acid combined and juxtaposed in an arrangement not existing in nature 
     “Complementary DNA (cDNA)” is a single-stranded DNA molecule that is formed from an mRNA template by the enzyme reverse transcriptase. Typically, a primer complementary to portions of mRNA is employed for the initiation of reverse transcription. Those skilled in the art also use the term “cDNA” to refer to a double-stranded DNA molecule consisting of such a single-stranded DNA molecule and its complementary DNA strand. The term “cDNA” also refers to a clone of a cDNA molecule synthesized from an RNA template. 
     A “promoter” is a nucleotide sequence that directs the transcription of a structural gene. Typically, a promoter is located in the 5′ non-coding region of a gene, proximal to the transcriptional start site of a structural gene. Sequence elements within promoters that function in the initiation of transcription are often characterized by consensus nucleotide sequences. These promoter elements include RNA polymerase binding sites, TATA sequences, CAAT sequences, differentiation-specific elements (DSEs; McGehee et al., Mol. Endocrinol., 7:551 (1993)), cyclic AMP response elements (CREs), serum response elements (SREs; Treisman, Seminars in Cancer Biol., 1:47 (1990)), glucocorticoid response elements (GREs), and binding sites for other transcription factors, such as CRE/ATF (O&#39;Reilly et al., J. Biol. Chem., 267:19938 (1992)), AP2 (Ye et al., J. Biol. Chem., 269:25728 (1994)), SP1, cAMP response element binding protein (CREB; Loeken, Gene Expr., 3:253 (1993)) and octamer factors (see, in general, Watson et al., eds., Molecular Biology of the Gene, 4th Edition, The Benjamin/Cummings Publishing Company, Inc. (1987), and Lemaigre et al., Biochem. J., 303:1 (1994)). If a promoter is an inducible promoter, then the rate of transcription increases in response to an inducing agent. In contrast, the rate of transcription is not regulated by an inducing agent if the promoter is a constitutive promoter. Repressible promoters are also known. 
     A “regulatory element” is a nucleotide sequence that modulates the activity of a core promoter. For example, a regulatory element may contain a nucleotide sequence that binds with cellular factors enabling transcription exclusively or preferentially in particular cells, tissues, or organelles. These types of regulatory elements are normally associated with genes that are expressed in a “cell-specific”, “tissue-specific”, or “organelle-specific” manner. 
     An “enhancer” is a type of regulatory element that can increase the efficiency of transcription, regardless of the distance or orientation of the enhancer relative to the start site of transcription. 
     “Heterologous DNA” refers to a DNA molecule, or a population of DNA molecules, that does not exist naturally within a given host cell. DNA molecules heterologous to a particular host cell may contain DNA derived from the host cell species (i.e., endogenous DNA) so long as that host DNA is combined with non-host DNA (i.e., exogenous DNA). For example, a DNA molecule containing a non-host DNA segment encoding a polypeptide operably linked to a host DNA segment comprising a transcription promoter is considered to be a heterologous DNA molecule. Conversely, a heterologous DNA molecule can comprise an endogenous gene operably linked with an exogenous promoter. As another illustration, a DNA molecule comprising a gene derived from a wild-type cell is considered to be heterologous DNA if that DNA molecule is introduced into a mutant cell that lacks the wild-type gene. 
     An “expression vector” is a nucleic acid molecule encoding a gene that is expressed in a host cell. Typically, an expression vector comprises a transcription promoter, a gene, and a transcription terminator. Gene expression is usually placed under the control of a promoter, and such a gene is said to be “operably linked to” the promoter. Similarly, a regulatory element and a core promoter are operably linked if the regulatory element modulates the activity of the core promoter. 
     A “recombinant host” is a cell that contains a heterologous nucleic acid molecule, such as a cloning vector or expression vector. In the present context, an example of a recombinant host is a cell that produces an antagonist of the present invention from an expression vector. In contrast, such an antagonist can be produced by a cell that is a “natural source” of said antagonist, and that lacks an expression vector. 
     The terms “amino-terminal” and “carboxyl-terminal” are used herein to denote positions within polypeptides. Where the context allows, these terms are used with reference to a particular sequence or portion of a polypeptide to denote proximity or relative position. For example, a certain sequence positioned carboxyl-terminal to a reference sequence within a polypeptide is located proximal to the carboxyl terminus of the reference sequence, but is not necessarily at the carboxyl terminus of the complete polypeptide. 
     A “fusion protein” is a hybrid protein expressed by a nucleic acid molecule comprising nucleotide sequences of at least two genes. For example, a fusion protein can comprise at least part of a IL-17RA polypeptide fused with a polypeptide that binds an affinity matrix. Such a fusion protein provides a means to isolate large quantities of IL-17A using affinity chromatography. 
     The term “receptor” denotes a cell-associated protein that binds to a bioactive molecule termed a “ligand.” This interaction mediates the effect of the ligand on the cell. Receptors can be membrane bound, cytosolic or nuclear; monomeric (e.g., thyroid stimulating hormone receptor, beta-adrenergic receptor) or multimeric (e.g., PDGF receptor, growth hormone receptor, IL-3 receptor, GM-CSF receptor, G-CSF receptor, erythropoietin receptor and IL-6 receptor). Membrane-bound receptors are characterized by a multi-domain structure comprising an extracellular ligand-binding domain and an intracellular effector domain that is typically involved in signal transduction. In certain membrane-bound receptors, the extracellular ligand-binding domain and the intracellular effector domain are located in separate polypeptides that comprise the complete functional receptor. In general, the binding of ligand to receptor results in a conformational change in the receptor that causes an interaction between the effector domain and other molecule(s) in the cell, which in turn leads to an alteration in the metabolism of the cell. Metabolic events that are often linked to receptor-ligand interactions include gene transcription, phosphorylation, dephosphorylation, increases in cyclic AMP production, mobilization of cellular calcium, mobilization of membrane lipids, cell adhesion, hydrolysis of inositol lipids and hydrolysis of phospholipids. 
     The term “expression” refers to the biosynthesis of a gene product. For example, in the case of a structural gene, expression involves transcription of the structural gene into mRNA and the translation of mRNA into one or more polypeptides. 
     The term “complement/anti-complement pair” denotes non-identical moieties that form a non-covalently associated, stable pair under appropriate conditions. For instance, biotin and avidin (or streptavidin) are prototypical members of a complement/anti-complement pair. Other exemplary complement/anti-complement pairs include receptor/ligand pairs, antibody/antigen (or hapten or epitope) pairs, sense/antisense polynucleotide pairs, and the like. Where subsequent dissociation of the complement/anti-complement pair is desirable, the complement/anti-complement pair preferably has a binding affinity of less than 10 9  M −1 . 
     A “detectable label” is a molecule or atom which can be conjugated to an antibody moiety to produce a molecule useful for diagnosis. Examples of detectable labels include chelators, photoactive agents, radioisotopes, fluorescent agents, paramagnetic ions, or other marker moieties. 
     The term “affinity tag” is used herein to denote a polypeptide segment that can be attached to a second polypeptide to provide for purification or detection of the second polypeptide or provide sites for attachment of the second polypeptide to a substrate. In principal, any peptide or protein for which an antibody or other specific binding agent is available can be used as an affinity tag. Affinity tags include a polyhistidine tract, protein A (Nilsson et al., EMBO J., 4:1075 (1985); Nilsson et al., Methods Enzymol., 198:3 (1991)), glutathione S transferase (Smith et al., Gene, 67:31 (1988)), Glu-Glu affinity tag (Grussenmeyer et al., Proc. Natl. Acad. Sci. USA, 82:7952 (1985)), substance P, FLAG® peptide (Hopp et al., Biotechnology, 6:1204 (1988)), streptavidin binding peptide, or other antigenic epitope or binding domain. See, in general, Ford et al., Protein Expression and Purification, 2:95 (1991). DNA molecules encoding affinity tags are available from commercial suppliers (e.g., Pharmacia Biotech, Piscataway, N.J.). 
     The term “acidic residue” refers to amino acid residues having sidechains comprising acidic groups. Exemplary acidic residues include D and E. 
     The term “amide residue” refers to amino acids having sidechains comprising amide derivatives of acidic groups. Exemplary residues include N and Q. 
     The term “aromatic residue” refers to amino acid residues having sidechains comprising aromatic groups. Exemplary aromatic residues include F, Y, and W. 
     The term “basic residue” refers to amino acid residues having sidechains comprising basic groups. Exemplary basic residues include H, K, and R. 
     The term “hydrophilic residue” refers to amino acid residues having sidechains comprising polar groups. Exemplary hydrophilic residues include C, S, T, N, and Q. 
     The term “non-functional residue” refers to amino acid residues having sidechains that lack acidic, basic, or aromatic groups. Exemplary non-functional amino acid residues include M, G, A, V, I, L and norleucine (Nle). 
     Both the EU index as in Kabat et al. (1991),  Sequences of Proteins of Immunological Interest,  5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD and AHo numbering schemes (Honegger and Plückthun (2001),  J Mol Biol.  8; 309(3): 657-70) can be used in the present invention. Amino acid positions and CDRs and FRs of a given antibody may be identified using either system. For example, EU heavy chain positions 39, 44, 183, 356, 357, 370, 392, 399, and 409 are equivalent to AHo heavy chain positions 46, 51, 230, 484, 485, 501, 528, 535, and 551, respectively. Similarly, EU light chain positions 38, 100, and 176 are equivalent to AHO light chain positions 46 141, and 230, respectively. Tables (i), (ii) and (iii) below demonstrate the equivalence between numbering positions for v1, v2, and v3 versions of charge positions to aid correct assembly of, for example, IgG-Fab bispecific antigen binding proteins. 
     
       
         
           
               
               
               
               
               
               
               
             
               
                   
                 TABLE (i)-v1 
               
               
                   
                   
               
               
                   
                 Chain 
                 Domain 
                 Mutation 
                 AHo # 
                 EU # 
                 Kabat # 
               
               
                   
                   
               
             
            
               
                   
                 LC-E 
                 Constant 
                 E 
                 230 
                 176 
                 176 
               
               
                   
                 LC-K 
                 Constant 
                 K 
                 230 
                 176 
                 176 
               
               
                   
                 HC-E 
                 CH1 
                 E 
                 230 
                 183 
                 188 
               
               
                   
                 HC-K 
                 CH1 
                 K 
                 230 
                 183 
                 188 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
             
               
                   
                 TABLE (ii)-v2 
               
               
                   
                   
               
               
                   
                 Chain 
                 Domain 
                 Mutation 
                 AHo # 
                 EU # 
                 Kabat # 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 LC-E 
                 Variable 
                 E 
                 46 
                 38 
                 38 
               
               
                   
                   
                 Constant 
                 E 
                 230 
                 176 
                 176 
               
               
                   
                 LC-K 
                 Variable 
                 K 
                 46 
                 38 
                 38 
               
               
                   
                   
                 Constant 
                 K 
                 230 
                 176 
                 176 
               
               
                   
                 HC-E 
                 Variable 
                 E 
                 46 
                 39 
                 39 
               
               
                   
                   
                 CH1 
                 E 
                 230 
                 183 
                 188 
               
               
                   
                 HC-K 
                 Variable 
                 K 
                 46 
                 39 
                 39 
               
               
                   
                   
                 CH1 
                 K 
                 230 
                 183 
                 188 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
             
               
                   
                 TABLE (iii)-v3 
               
               
                   
                   
               
               
                   
                 Chain 
                 Domain 
                 Mutation 
                 AHo # 
                 EU # 
                 Kabat # 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 LC-E 
                 Variable 
                 E 
                 141 
                 100 
                 100 
               
               
                   
                   
                 Constant 
                 E 
                 230 
                 176 
                 176 
               
               
                   
                 LC-K 
                 Variable 
                 K 
                 141 
                 100 
                 100 
               
               
                   
                   
                 Constant 
                 K 
                 230 
                 176 
                 176 
               
               
                   
                 HC-E 
                 Variable 
                 E 
                 51 
                 44 
                 44 
               
               
                   
                   
                 CH1 
                 E 
                 230 
                 183 
                 188 
               
               
                   
                 HC-K 
                 Variable 
                 K 
                 51 
                 44 
                 44 
               
               
                   
                   
                 CH1 
                 K 
                 230 
                 183 
                 188 
               
               
                   
                   
               
            
           
         
       
     
     Bispecific Antigen Binding Protein Formats of the Invention 
     One aspect of the invention concerns novel TL1A-specific antigen binding proteins and antibodies. Such antibodies are useful to treat conditions known in the art and described hereinafter that are amenable to treatment by inhibition of TL1A biological activity. 
     Another aspect of the invention concerns bispecific antigen binding proteins in which one light chain-heavy chain pair specifically binds TL1A and another light chain-heavy chain pair binds TNF-α. Such bispecific antigen binding proteins can be whole antibodies (see  FIG. 1 ) or F(ab′) 2  fragments. In this aspect of the invention, the TL1A binding entity is monovalent for TL1A and the TNF-α binding entity is monovalent for TNF-α. 
     In another bispecific antigen binding protein embodiment, the TL1A binding entity and the TNF-α binding entity are arranged in an overall structure referred to herein as IgG-scFv. In this embodiment, a first binding entity has the structure of an antibody—i.e., two pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. A second binding entity comprises the structure of a pair of Fv units—i.e., each member of the pair has a variable domain from a heavy chain and a variable domain from a light chain linked in tandem as a single chain. In the IgG-scFv configuration, each Fv unit is covalently bound to the C-terminus of a heavy chain constant domain (Fc) of the first binding entity (see  FIG. 2 ). Each Fv unit can be linked to the Fc domain of the first binding entity directly or through a linker. In this embodiment of the invention, each binding entity is bivalent for its target antigen. In a preferred embodiment, the TL1A binding entity has the structure of an antibody and the TNF-α binding entity has the structure of a pair of Fv units. 
     The invention further concerns bispecific antibodies with mutations to aid correct heavy-heavy and heavy-light chain pairing. Such mutations are described in U.S. Pat. No. 8,592,562; WO 2009/089004; WO 2006/106905; WO 2014/4082179; WO 2014/081955; Speiss et al.,  Mol. Immunol.  (2015), http://dx.doi.org/10.1016/j.molimm.2015.01.003; each of which is hereby incorporated by reference. 
     The invention further concerns a TL1A binding entity and a TNF-α binding entity arranged in other bispecific antigen binding protein formats known in the art. Specifically, the invention concerns the TL1A and TNF-α binding entities in the formats described in: WO 2014/159 725; WO 2013/041687; U.S. Pat. Nos. 8,945,553; 8,945,553; 8,258,268; US Pat. App. No. 2012/0195900; International patent application WO 2012/088 302; US Prov. App. 62/218,977; Fischer and Leger (2007),  Pathobiology  74:3-14; van Spriel et al. (2000),  Immunology Today  21(8): 391-7; Kufer et al. (2004),  Trends in Biotechnology  22(5): 238-44; Byrne et al. (2013),  Trends in Biotechnology  31(11): 621-31; Muller and Kontermann (2010),  Biodrugs  24(2): 89-98; Chames and Baty (2009), http://dx.doi.org/10.4161/mabs.1.6.10015; Kontermann (2012), http://dx.doi.org/10.4161/mabs.4.2.19000; Holliger et al. (1993),  Proc. Natl. Acad. Sci. USA  90: 6444-8; Speiss et al.,  Mol. Immunol.  (2015), http://dx.doi.org/10.1016/j.molimm.2015.01.003; WO 2009/089004, published 16 Jul. 2009; Holliger et al. (1993),  Proc. Natl. Acad. Sci. USA  90: 6444-8; Speiss et al.,  Mol. Immunol.  (2015), http://dx.doi.org/10.1016/j.molimm.2015.01.003; Ridgway et al, (1996),  Protein Eng.  9: 617; Gunasekaran et al (2010),  J. Biol. Chem.  285:19637; Davis (2010),  Protein Eng. Des.  &amp;  Sel.  23:195; DiGiammarino et al. (2012),  Methods Mol. Biol.  899:145, 2012); Lindhofer et al. (1995),  J. Immunol.  155: 219: Schaefer et al. (2011),  Proc. Natl. Acad. Sci. USA,  108: 11187; Regula et al, US Patent Application No: 2010/0322934; Bostrom et al. (2009),  Science  323:1610; US Pat. App. 2011/0076722; Rossi et al. (2008),  Cancer Res.  68:8384; each of which is hereby incorporated by reference. 
     Preferred Embodiments 
     The amino acid sequences of the antigen binding proteins and binding entities are preferably based upon the sequences of human and/or humanized monoclonal antibodies against TL1A and TNF-α. The invention also comprises sequences having at least 90%, at least 95%, or at least 99% sequence identity to the preferred sequences set forth hereinafter. The amino acid sequences shown in the following tables are preferred for the antigen binding proteins of this invention, including bispecific antigen binding proteins in any format. 
     TL1A-Specific Antigen Binding Proteins 
     TL1A-specific antigen binding proteins and antigen binding entities preferably comprise the complementarity determining region (CDR) sequences derived from preferred TL1A antibodies disclosed herein. Table A shows the preferred CDR sequences, alongside the preferred antibodies from which they were derived. Throughout, LCDR1, LCDR2, and LCDR3 refer to the light chain CDRs; HCDR1, HCDR2, and HCDR3 to the heavy chain CDRs, Throughout, the sequence identification number (SEQ ID NO) for each sequence appears in parentheses after the sequence in the table. 
     
       
         
           
               
             
               
                 TABLE A 
               
             
            
               
                   
               
               
                 Preferred TL1A-binding CDR sequences 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Source 
                 LCDR1 
                 LCDR2 
                 LCDR3 
                 HCDR1 
                 HCDR2 
                 HCDR3 
               
               
                 Antibody 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
               
               
                 designation 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
               
               
                   
               
               
                 3C6 
                 RTSQDIRD 
                 DASSLQS 
                 LQHNSYPP 
                 SYGMH 
                 VISYDGNN 
                 PTVTLYYY 
               
               
                   
                 DLG (98) 
                 (100) 
                 1(102) 
                 (164) 
                 KLYTDSVK 
                 YGMDV 
               
               
                   
                   
                   
                   
                   
                 G (166) 
                 (168) 
               
               
                   
               
               
                 2G11 
                 RASQSINN 
                 ATSSLQS 
                 QQSYSTPR 
                 SYFWS 
                 YIYYSGST 
                 EIGSYYGF 
               
               
                   
                 YLN (110) 
                 (106) 
                 T (108) 
                 (170) 
                 NYNPSLKS 
                 DY (174) 
               
               
                   
                   
                   
                   
                   
                 (172) 
                   
               
               
                   
               
               
                 9C8 
                 RASQSINN 
                 AASSLQS 
                 QQSYSTPR 
                 SYFWS 
                 YIYYSGNT 
                 ETGSYYGF 
               
               
                   
                 YLN (110) 
                 (112) 
                 T (108) 
                 (170) 
                 KYNPSLKS 
                 DY (180) 
               
               
                   
                   
                   
                   
                   
                 (178) 
                   
               
               
                   
               
               
                 23B3 
                 KSSQSVLY 
                 WASTRES 
                 QQYYKTPL 
                 TNSVAWN 
                 RTYYRSK 
                 EDGDSYYR 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 T (120) 
                 (182) 
                 WYNDYAV 
                 YGMDV 
               
               
                   
                 V (116) 
                   
                   
                   
                 SVKS (184) 
                 (186) 
               
               
                   
               
               
                 23B3 VH4 
                 RSSQSVLY 
                 WASTRES 
                 QQYYKTPL 
                 TNSVAWN 
                 RTYYRSK 
                 EDGDSYYR 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 T (120) 
                 (182) 
                 WYNDYAV 
                 YGMDV 
               
               
                   
                 V (128) 
                   
                   
                   
                 SLKS (196) 
                 (186) 
               
               
                   
               
               
                 23B3 VH3 
                 RSSQSVLY 
                 WASTRES 
                 QQYYKTPL 
                 TNSVAWN 
                 RTYYRSK 
                 EDGDSYYR 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 T (120) 
                 (182) 
                 WYNDYAV 
                 YGMDV 
               
               
                   
                 V (128) 
                   
                   
                   
                 SVKG (202) 
                 (186) 
               
               
                   
               
               
                 3B3 
                 RASQSVRS 
                 GASSRAT 
                 QQYGSSPT 
                 GYYWN 
                 EINHAGNT 
                 GYCRSTTC 
               
               
                   
                 SYLA (122) 
                 (124) 
                 (126) 
                 (188) 
                 NYNPSLKS 
                 YFDY (192) 
               
               
                   
                   
                   
                   
                   
                 (190) 
                   
               
               
                   
               
               
                 5G4 
                 RASQSVRS 
                 GASSRAT 
                 QQYGSSPT 
                 GYYWN 
                 EINHSGITN 
                 GYCRSTTC 
               
               
                   
                 SYLA (122) 
                 (124) 
                 (126) 
                 (188) 
                 YNPSLKS 
                 YFDY (192) 
               
               
                   
                   
                   
                   
                   
                 (483) 
                   
               
               
                   
               
               
                 17E9 
                 RASQGISN 
                 AASSLQS 
                 LQHNSYPP 
                 SYGMH 
                 VMSYDGN 
                 DETETLYY 
               
               
                   
                 DLA (469) 
                 (112) 
                 T (102) 
                 (164) 
                 NKLYADSV 
                 YYGIDV 
               
               
                   
                   
                   
                   
                   
                 KG (485) 
                 (489) 
               
               
                   
               
               
                 53D3 
                 RSSQSLLY 
                 LGSSRAS 
                 MQPLQTPL 
                 TYYMS 
                 SISSSSSFI 
                 DRIAAPGT 
               
               
                   
                 SNGYNYLD 
                 (685) 
                 T (687) 
                 (777) 
                 YYADSVKG 
                 YYYYGMD 
               
               
                   
                 (683) 
                   
                   
                   
                 (779) 
                 V (781) 
               
               
                   
               
               
                 54E5 
                 KSSQNILYS 
                 WASTRES 
                 QQYYSIPW 
                 TYGMH 
                 VIWYDGSN 
                 EERDSYYH 
               
               
                   
                 SNNKNYLA 
                 (118) 
                 T (693) 
                 (783) 
                 KDYADSVK 
                 YGMDV 
               
               
                   
                 (689) 
                   
                   
                   
                 G (785) 
                 (787) 
               
               
                   
               
               
                 56E1 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSIPW 
                 SYGMH 
                 VIWYDGSN 
                 EERDSYYH 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 T (693) 
                 (164) 
                 KDYADSVK 
                 YGMDV 
               
               
                   
                 A (235) 
                   
                   
                   
                 G (785) 
                 (787) 
               
               
                   
               
               
                 57A8 
                 TGSSSNIG 
                 GNNNRPS 
                 QSYDSSLS 
                 SYVMS 
                 VISGSGGS 
                 GGTNYYYY 
               
               
                   
                 AGYNVH 
                 (699) 
                 GWV (701) 
                 (792) 
                 TYYADSVK 
                 YSGMDV 
               
               
                   
                 (697) 
                   
                   
                   
                 G (794) 
                 (796) 
               
               
                   
               
               
                 58G5 
                 TGSSSNIG 
                 GNSHRPS 
                 QSYDSSLS 
                 NYAMS 
                 VISGRGGS 
                 DGYSSAW 
               
               
                   
                 AGYDVH 
                 (705) 
                 GYV (707) 
                 (798) 
                 TYYADSVK 
                 YFDY (802) 
               
               
                   
                 (146) 
                   
                   
                   
                 G (800) 
                   
               
               
                   
               
               
                 60G11 
                 TGSGSNIG 
                 GNSHRPS 
                 QSYDSSLS 
                 NYAMN 
                 VISGRGGS 
                 DGYSSAW 
               
               
                   
                 AGYDVH 
                 (705) 
                 GYV (707) 
                 (804) 
                 TYYADSVK 
                 FFDY (807) 
               
               
                   
                 (709) 
                   
                   
                   
                 G (800) 
                   
               
               
                   
               
               
                 73C2 
                 RASQSFSS 
                 AASSLQS 
                 QQSYSTPR 
                 SSSATWN 
                 RTYQRSK 
                 EVVAGPR 
               
               
                   
                 YLN (713) 
                 (112) 
                 S (717) 
                 (809) 
                 WNNDYAV 
                 WFDP (813) 
               
               
                   
                   
                   
                   
                   
                 SVKS (811) 
                   
               
               
                   
               
               
                 76A4 
                 RASQSVTS 
                 TASSLQS 
                 QQSYSTPR 
                 SNSATWN 
                 RTYYRSKL 
                 EVVAGPR 
               
               
                   
                 YLN (719) 
                 (721) 
                 S (717) 
                 (815) 
                 YNDYAVSV 
                 WFDP (813) 
               
               
                   
                   
                   
                   
                   
                 KS (817) 
                   
               
               
                   
               
               
                 77D12 
                 SESNSDIG 
                 SNNKRPS 
                 ATWDDNL 
                 GFYMH 
                 WINPDSGG 
                 EGIAVALTY 
               
               
                   
                 TNAVN 
                 (725) 
                 NGPL (727) 
                 (819) 
                 TNYAQKFQ 
                 (823) 
               
               
                   
                 (723) 
                   
                   
                   
                 G (821) 
                   
               
               
                   
               
               
                 87H11 
                 RASQSISS 
                 VASSLQS 
                 QQSYSNP 
                 GYYWS 
                 EINHSGRT 
                 DSGWHFS 
               
               
                   
                 YLN (229) 
                 (731) 
                 QECS (733) 
                 (265) 
                 NYNPSLKS 
                 FDI (829) 
               
               
                   
                   
                   
                   
                   
                 (827) 
                   
               
               
                   
               
               
                 88H9 
                 RASQGIRN 
                 AASGLQG 
                 LQHNSYPT 
                 SYGMH 
                 VIWFDGSN 
                 ERWFGELL 
               
               
                   
                 DLG (226) 
                 (737) 
                 WT (739) 
                 (164) 
                 EYYADSVK 
                 DY (834) 
               
               
                   
                   
                   
                   
                   
                 G (832) 
                   
               
               
                   
               
               
                 89H9 
                 RASQSISY 
                 AASSLQS 
                 QQSYSSIT 
                 SYAMS 
                 GISGGGGS 
                 EMAGAFDI 
               
               
                   
                 YLN (741) 
                 (112) 
                 (745) 
                 (836) 
                 TYYADSVK 
                 (840) 
               
               
                   
                   
                   
                   
                   
                 G (838) 
                   
               
               
                   
               
               
                 91D7 
                 RASQNISS 
                 TASSLQS 
                 QQSYSNPP 
                 GYYWS 
                 EINPVGRT 
                 DNGWHYA 
               
               
                   
                 YLN (747) 
                 (749) 
                 ESS (751) 
                 (265) 
                 NYKPSLKS 
                 FDI (845) 
               
               
                   
                   
                   
                   
                   
                 (843) 
                   
               
               
                   
               
               
                 91F10_LC1 
                 RSSQSLVY 
                 KVSNWDS 
                 MQGTHWP 
                 AYYMH 
                 WINPNSGG 
                 GGSWEGF 
               
               
                   
                 SDGNTYLN 
                 (755) 
                 (757) 
                 (847) 
                 TNYAQKFR 
                 DY (851) 
               
               
                   
                 (753) 
                   
                   
                   
                 G (849) 
                   
               
               
                   
               
               
                 91F10-LC2 
                 RASQSISS 
                 AASRLQS 
                 QQSDSTPI 
                 AYYMH 
                 WINPNSGG 
                 GGSWEGF 
               
               
                   
                 YLN (229) 
                 (761) 
                 T (763) 
                 (847) 
                 TNYAQKFR 
                 DY (851) 
               
               
                   
                   
                   
                   
                   
                 G (849) 
                   
               
               
                   
               
               
                 91G8 
                 RASQSISS 
                 AASSLQS 
                 QQSFSTIT 
                 SYAMS 
                 GISGSGGS 
                 EVAGAFDI 
               
               
                   
                 YLN (229) 
                 (112) 
                 (767) 
                 (836) 
                 RYYADSVK 
                 (855) 
               
               
                   
                   
                   
                   
                   
                 G (853) 
                   
               
               
                   
               
               
                 92D3 
                 RASQSISS 
                 GASRLQS 
                 QQSDTTPI 
                 GYYMH 
                 WIIPNSGG 
                 GSSWEGF 
               
               
                   
                 YLN (229) 
                 (769) 
                 T (771) 
                 (857) 
                 TNYAQKFQ 
                 DY (861) 
               
               
                   
                   
                   
                   
                   
                 G (859) 
                   
               
               
                   
               
               
                 92E5 
                 RASQSISH 
                 AASSLQS 
                 QQSFSSIT 
                 SYAMS 
                 GISGRGGS 
                 EVAGAFDI 
               
               
                   
                 YLN (773) 
                 (112) 
                 (775) 
                 (836) 
                 TYYADSVK 
                 (855) 
               
               
                   
                   
                   
                   
                   
                 G (864) 
               
               
                   
               
            
           
         
       
     
     Each of the foregoing sequences is encoded by the nucleic acid sequence immediately preceding it in the Sequence Listing. Throughout, the sequences from antibodies 9C8 and 3B3 are preferred. 
     In the antigen binding proteins of this invention, it is preferable to avoid isomerization sites. The two-amino acid sequences DG, DH, DS, and DT are known isomerization sites. The present invention relates also to antigen binding proteins in which the source antibody sequences, including the sequences of CDRs, are modified to eliminate isomerization sites. With that consideration, the invention relates to antigen binding proteins wherein HCDR2 is a modified form of the HCDR2 from source antibody 3C6 having the sequence VISYDXNNKLYTDXVKG (SEQ ID NO: 204) wherein each X is independently a residue other than G, H, S, or T (i.e., A, C, D, E, F, I, K, L, M, N, P, Q, R, V, W, or Y), with A preferred. The invention further relates to antigen binding proteins wherein HCDR2 is a modified form of the HCDR2 from source antibody 3C6 in which the acidic residue D is replaced with E so as to remove the isomerization sites, resulting in the sequence VISYEGNNKLYTESVKG (SEQ ID NO: 678). The invention further relates to antigen binding proteins wherein HCDR3 is a modified form of HCDR3 from source antibody 23B3 having the sequence EDGDXYYRYGMDV (SEQ ID NO: 676). The invention further relates to antigen binding proteins wherein the isomerization site of HCDR3 from source antibody 23B3 is removed by replacing the acidic residue D with E, resulting in the sequence EEGESYYRYGMDV (SEQ ID NO: 657). 
     In the antigen binding proteins of this invention, it is preferable to avoid deamidation sites. The two-amino acid sequences NG, NH, NS, and NT are known deamidation sites. The present invention relates also to antigen binding proteins in which the source antibody sequences, including the sequences of CDRs, are modified to eliminate deamidation sites. One way to eliminate the deamidation sites is to replace the second amino acid residue in the sites NG, NH, NS, and NT with a residue other than G, H, S, or T (i.e., A, C, D, E, F, I, K, L, M, N, P, Q, R, V, W, or Y). A preferred way to eliminate deamidation sites is to replace the N in NG, NH, NS and NT with Q. With that consideration, the invention relates to antigen binding proteins having the following sequences in addition to those listed in Table A: 
     LCDR3 may have the sequence LQHQSYPPT (SEQ ID NO: 631) based on modification of the sequence from the source antibody 3C6; 
     HCDR1 may have the sequences TQSVAWN (SEQ ID NO: 632) based on modification of the sequence from the source antibody 23B3 VH4; 
     HCDR2 may have the sequences YIYYSGQTKYNPSLKS (SEQ ID NO: 633) or EIQHAGQTNYNPSLKS (SEQ ID NO: 677) based on modification of the sequences from the source antibodies 9C8 and 3B3, respectively. 
     Antigen binding proteins of this invention may further result from substitution of an acidic residue for another acidic residue, an amide residue for another amide residue, and likewise for aromatic residues, basic residues, hydrophilic residues, non-functional residues, neutral polar residues, and polar hydrophobic residues. Such substitutions of non-functional residues of the source antibody CDRs results in the sequences of the invention shown in Table B, wherein each X is independently M, G, A, V, I, or L. 
     
       
         
           
               
             
               
                 TABLE B 
               
             
            
               
                   
               
               
                 TL1A-binding CDR sequences with substitutions 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Source 
                 LCDR1 
                 LCDR2 
                 LCDR3 
                 HCDR1 
                 HCDR2 
                 HCDR3 
               
               
                 Antibody 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
               
               
                 designation 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
               
               
                   
               
               
                 3C6 
                 RTSQDXRD 
                 DXSSLQS 
                 XQHQSYPP 
                 SYXXH 
                 XXSYDXNN 
                 PTXTXYYY 
               
               
                   
                 DLX (635) 
                 (636) 
                 T (637), 
                 (639) 
                 KXYTDXXK 
                 YXXDV 
               
               
                   
                   
                   
                 XQHNXYPP 
                   
                 X (640), 
                 (641) 
               
               
                   
                   
                   
                 T (638), 
                   
                 VISYDXNN 
                   
               
               
                   
                   
                   
                 LQHQSYPP 
                   
                 KLYTDXVK 
                   
               
               
                   
                   
                   
                 T (631), 
                   
                 G (204), 
                   
               
               
                   
                   
                   
                   
                   
                 VISYEGNN 
                   
               
               
                   
                   
                   
                   
                   
                 KLYTESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G(678), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSYEXNN 
                   
               
               
                   
                   
                   
                   
                   
                 KLYTESXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (679) 
                   
               
               
                   
               
               
                 2G11 
                 RXSQSXNN 
                 XTSSXQS 
                 QQSYSTPR 
                 SYFWS 
                 YXYYSXST 
                 EXXSYYXF 
               
               
                   
                 YLN (642) 
                 (643) 
                 T (108) 
                 (170) 
                 NYNPSXKS 
                 DY (645) 
               
               
                   
                   
                   
                   
                   
                 (644) 
                   
               
               
                   
               
               
                 9C8 
                 RXSQSXNN 
                 XXSSXQS 
                 QQSYSTPR 
                 SYFWS 
                 YXYYSXQT 
                 ETXSYYXF 
               
               
                   
                 YXN (646) 
                 (647) 
                 T (108) 
                 (170) 
                 KYNPSXKS 
                 DY (650) 
               
               
                   
                   
                   
                   
                   
                 (648), 
                   
               
               
                   
                   
                   
                   
                   
                 YXYYSXNX 
                   
               
               
                   
                   
                   
                   
                   
                 KYNPSXKS 
                   
               
               
                   
                   
                   
                   
                   
                 (649), 
                   
               
               
                   
                   
                   
                   
                   
                 YIYYSGQT 
                   
               
               
                   
                   
                   
                   
                   
                 KYNPSLKS 
                   
               
               
                   
                   
                   
                   
                   
                 (633) 
                   
               
               
                   
               
               
                 23B3 
                 KSSQSVXY 
                 WXSTRES 
                 QQYYKTPX 
                 TQSXXWN 
                 RTYYRSK 
                 EDXDSYYR 
               
               
                   
                 SSNNKNYX 
                 (652) 
                 T (653) 
                 (654), 
                 WYNDYXV 
                 YXXDV 
               
               
                   
                 X (651) 
                   
                   
                 TNXXXWN 
                 SXKS (656) 
                 (657), 
               
               
                   
                   
                   
                   
                 (655), 
                   
                 EDGDXYYR 
               
               
                   
                   
                   
                   
                 TQSVAWN 
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                 (632) 
                   
                 (676), 
               
               
                   
                   
                   
                   
                   
                   
                 EEGESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                   
                   
                 (680), 
               
               
                   
                   
                   
                   
                   
                   
                 EEXESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YXXDX 
               
               
                   
                   
                   
                   
                   
                   
                 (681) 
               
               
                   
               
               
                 23B3 VH4 
                 RSSQSVXY 
                 WXSTRES 
                 QQYYKTPX 
                 TQSXXWN 
                 RTYYRSK 
                 EDXDSYYR 
               
               
                   
                 SSNNKNYX 
                 (652) 
                 T (653) 
                 (654), 
                 WYNDYXX 
                 YXXDV 
               
               
                   
                 X (658) 
                   
                   
                 TNXXXWN 
                 SXKS (659) 
                 (657), 
               
               
                   
                   
                   
                   
                 (655), 
                   
                 EDGDXYYR 
               
               
                   
                   
                   
                   
                 TQSVAWN 
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                 (632) 
                   
                 (676), 
               
               
                   
                   
                   
                   
                   
                   
                 EEGESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                   
                   
                 (680), 
               
               
                   
                   
                   
                   
                   
                   
                 EEXESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YXXDX 
               
               
                   
                   
                   
                   
                   
                   
                 (681) 
               
               
                   
               
               
                 23B3 VH3 
                 RSSQSVXY 
                 WXSTRES 
                 QQYYKTPX 
                 TQSXXWN 
                 RTYYRSK 
                 EDXDSYYR 
               
               
                   
                 SSNNKNYX 
                 (652) 
                 T (653) 
                 (654), 
                 WYNDYXX 
                 YXXDV 
               
               
                   
                 X (658) 
                   
                   
                 TNXXXWN 
                 SVKX (660) 
                 (657), 
               
               
                   
                   
                   
                   
                 (655), 
                   
                 EDGDXYYR 
               
               
                   
                   
                   
                   
                 TQSVAWN 
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                 (632) 
                   
                 (676), 
               
               
                   
                   
                   
                   
                   
                   
                 EEGESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YGMDV 
               
               
                   
                   
                   
                   
                   
                   
                 (680), 
               
               
                   
                   
                   
                   
                   
                   
                 EEXESYYR 
               
               
                   
                   
                   
                   
                   
                   
                 YXXDX 
               
               
                   
                   
                   
                   
                   
                   
                 (681) 
               
               
                   
               
               
                 3B3 
                 RXSQSVRS 
                 XXSSRXT 
                 QQYXSSPT 
                 XYYWN 
                 EXQHXXQT 
                 XYCRSTTC 
               
               
                   
                 SYXX (661) 
                 (662) 
                 (663) 
                 (664) 
                 NYNPSXKS 
                 YFDY (667) 
               
               
                   
                   
                   
                   
                   
                 (665), 
                   
               
               
                   
                   
                   
                   
                   
                 EXNHXXNT 
                   
               
               
                   
                   
                   
                   
                   
                 NYNPSXKS 
                   
               
               
                   
                   
                   
                   
                   
                 (666), 
                   
               
               
                   
                   
                   
                   
                   
                 EIQHAGQT 
                   
               
               
                   
                   
                   
                   
                   
                 NYNPSLKS 
                   
               
               
                   
                   
                   
                   
                   
                 (677) 
                   
               
               
                   
               
               
                 5G4 
                 RXSQSXRS 
                 XXSSRXT 
                 QQYXSSPT 
                 XYYWN 
                 EXNXSXXT 
                 XYCRSTTC 
               
               
                   
                 SYXX (668) 
                 (662) 
                 (663) 
                 (664) 
                 NYNPSXKS 
                 YFDY (667) 
               
               
                   
                   
                   
                   
                   
                 (114), 
                   
               
               
                   
                   
                   
                   
                   
                 EIQHSGITN 
                   
               
               
                   
                   
                   
                   
                   
                 YNPSLKS 
                   
               
               
                   
                   
                   
                   
                   
                 (227), 
                   
               
               
                   
                   
                   
                   
                   
                 XQHSXXTN 
                   
               
               
                   
                   
                   
                   
                   
                 YNPSXKS 
                   
               
               
                   
                   
                   
                   
                   
                 (230) 
                   
               
               
                   
               
               
                 17E9 
                 RXSQXXSN 
                 XXSSXQS 
                 XQHQSYPP 
                 SYXXH 
                 VMSYDXN 
                 DETETXYY 
               
               
                   
                 DX (646) 
                 (647) 
                 T (637), 
                 (639) 
                 NKLYADXV 
                 YYXXDX 
               
               
                   
                   
                   
                 XQHNSYPP 
                   
                 KG (691), 
                 (671) 
               
               
                   
                   
                   
                 T (233) 
                   
                 VMSYEGN 
                   
               
               
                   
                   
                   
                   
                   
                 NKLYAESV 
                   
               
               
                   
                   
                   
                   
                   
                 KG (715), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSYDXNN 
                   
               
               
                   
                   
                   
                   
                   
                 KXYXDSXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (669), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSYEXNN 
                   
               
               
                   
                   
                   
                   
                   
                 KXYXESXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (711) 
                   
               
               
                   
               
               
                 53D3 
                 RSSQSXXY 
                 XXSSRXS 
                 XQPXQTPX 
                 TYYXS 
                 SISSSSSFI 
                 DRXXXPXT 
               
               
                   
                 SNXYNYXD 
                 (789) 
                 T (940) 
                 (955) 
                 YYADXVKG 
                 YYYYXXDX 
               
               
                   
                 (759), 
                   
                   
                   
                 (971), 
                 (976) 
               
               
                   
                 RSSQSLLY 
                   
                   
                   
                 SISSSSSFI 
                   
               
               
                   
                 SQGYNYLD 
                   
                   
                   
                 YYAESVKG 
                   
               
               
                   
                 (743), 
                   
                   
                   
                 (972), 
                   
               
               
                   
                 RSSQSXXY 
                   
                   
                   
                 SXSSSSSF 
                   
               
               
                   
                 SQXYNYXD 
                   
                   
                   
                 XYYXDSXK 
                   
               
               
                   
                 (765) 
                   
                   
                   
                 X (973), 
                   
               
               
                   
                   
                   
                   
                   
                 SXSSSSSF 
                   
               
               
                   
                   
                   
                   
                   
                 XYYXESXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (974), 
                   
               
               
                   
                   
                   
                   
                   
                 SXSSSSSF 
                   
               
               
                   
                   
                   
                   
                   
                 XYYXDXXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (975) 
                   
               
               
                   
               
               
                 54E5 
                 KSSQNXLY 
                 WXSTRES 
                 QQYYSXP 
                 TYXXH 
                 VIWYDXSN 
                 EERDSYYH 
               
               
                   
                 SSNNKNYX 
                 (652) 
                 WT (978) 
                 (979) 
                 KDYADXVK 
                 YXXDX 
               
               
                   
                 X (977) 
                   
                   
                   
                 G (980), 
                 (985) 
               
               
                   
                   
                   
                   
                   
                 VIWYEGSN 
                   
               
               
                   
                   
                   
                   
                   
                 KDYAESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G (981), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYDXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXDSX 
                   
               
               
                   
                   
                   
                   
                   
                 KX (982), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYDXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXDXV 
                   
               
               
                   
                   
                   
                   
                   
                 KG (983), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYEXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXESX 
                   
               
               
                   
                   
                   
                   
                   
                 KG (984) 
                   
               
               
                   
               
               
                 56E1 
                 KSSQSXXY 
                 WXSTRES 
                 QQYYSXP 
                 SYXXH 
                 VIWYDXSN 
                 EERDSYYH 
               
               
                   
                 SSNNKNYX 
                 (652) 
                 WT (978) 
                 (639) 
                 KDYADXVK 
                 YXXDX 
               
               
                   
                 X (986) 
                   
                   
                   
                 G (980), 
                 (985) 
               
               
                   
                   
                   
                   
                   
                 VIWYEGSN 
                   
               
               
                   
                   
                   
                   
                   
                 KDYAESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G (981), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYDXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXDSX 
                   
               
               
                   
                   
                   
                   
                   
                 KX (982), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYDXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXDXV 
                   
               
               
                   
                   
                   
                   
                   
                 KG (983), 
                   
               
               
                   
                   
                   
                   
                   
                 XXWYEXS 
                   
               
               
                   
                   
                   
                   
                   
                 NKDYXESX 
                   
               
               
                   
                   
                   
                   
                   
                 KG (984) 
                   
               
               
                   
               
               
                 57A8 
                 TXSSSNXX 
                 XNNNRPS 
                 QSYDXSLS 
                 SYXXS 
                 VISGSGGS 
                 XXTNYYYY 
               
               
                   
                 XXYNXH 
                 (988) 
                 GWV (989), 
                 (994) 
                 TYYADXVK 
                 YSXXDX 
               
               
                   
                 (987) 
                   
                 QSYESSLS 
                   
                 G (995), 
                 (1000) 
               
               
                   
                   
                   
                 GWV (990), 
                   
                 VISGSGGS 
                   
               
               
                   
                   
                   
                 QSYDSSXS 
                   
                 TYYAESVK 
                   
               
               
                   
                   
                   
                 XWX (991), 
                   
                 G (996), 
                   
               
               
                   
                   
                   
                 QSYDXSXS 
                   
                 XXSXSXXS 
                   
               
               
                   
                   
                   
                 XWV (992), 
                   
                 TYYXDSXK 
                   
               
               
                   
                   
                   
                 QSYESSXS 
                   
                 X 
                   
               
               
                   
                   
                   
                 XWV (993) 
                   
                 (997), XXSX 
                   
               
               
                   
                   
                   
                   
                   
                 SXXSTYYX 
                   
               
               
                   
                   
                   
                   
                   
                 DXXKX 
                   
               
               
                   
                   
                   
                   
                   
                 (998), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSXSXXS 
                   
               
               
                   
                   
                   
                   
                   
                 TYYXESXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (999) 
                   
               
               
                   
               
               
                 58G5 
                 TXSSSNXX 
                 XNXHRPS 
                 QSYDXSLS 
                 NYXXS 
                 XXSXRXXS 
                 DXYSSAW 
               
               
                   
                 XXYDXH 
                 (1002), 
                 GYV 
                 (1010) 
                 TYYXDSXK 
                 YFDY 
               
               
                   
                 (1001) 
                 GQSHRPS 
                 (1005), QSY 
                   
                 X (1011) 
                 (1012), 
               
               
                   
                   
                 (1003), 
                 ESSLSGYV 
                   
                   
                 EGYSSAW 
               
               
                   
                   
                 XQSHRPS 
                 (1006), 
                   
                   
                 YFDY 
               
               
                   
                   
                 (1004) 
                 QSYDSSXS 
                   
                   
                 (1013), 
               
               
                   
                   
                   
                 XYX (1007), 
                   
                   
                 DXYSSXW 
               
               
                   
                   
                   
                 QSYDXSXS 
                   
                   
                 YFDY 
               
               
                   
                   
                   
                 XYX (1008), 
                   
                   
                 (1014), 
               
               
                   
                   
                   
                 QSYESSXS 
                   
                   
                 EXYSSXWY 
               
               
                   
                   
                   
                 XYX (1009) 
                   
                   
                 FDY (1015) 
               
               
                   
               
               
                 60G11 
                 TXSXSNXX 
                 XNXHRPS 
                 QSYDSSXS 
                 NYXXN 
                 XXSXRXXS 
                 DXYSSAWF 
               
               
                   
                 XXYDXH 
                 (1002), 
                 XYX (1007) 
                 (1017) 
                 TYYXDSXK 
                 FDY (1018), 
               
               
                   
                 (1016) 
                 GQSHRPS 
                   
                   
                 X (1011) 
                 EGYSSAW 
               
               
                   
                   
                 (1003), 
                   
                   
                   
                 FFDY 
               
               
                   
                   
                 XQSHRPS 
                   
                   
                   
                 (1019), 
               
               
                   
                   
                 (1004) 
                   
                   
                   
                 DXYSSXWF 
               
               
                   
                   
                   
                   
                   
                   
                 FDY (1020) 
               
               
                   
               
               
                 73C2 
                 RXSQSFSS 
                 XXSSXQS 
                 [none] 
                 SSSXTWN 
                 RTYQRSK 
                 EXXXXPR 
               
               
                   
                 YXN (1021) 
                 (647) 
                   
                 (1022) 
                 WNNDYXX 
                 WFDP 
               
               
                   
                   
                   
                   
                   
                 SXKS 
                 (1024) 
               
               
                   
                   
                   
                   
                   
                 (1023) 
                   
               
               
                   
               
               
                 76A4 
                 RXSQSXTS 
                 TXSSXQS 
                 [none] 
                 SNSXTWN 
                 RTYYRSKX 
                 EXXXXPR 
               
               
                   
                 YXN (1025) 
                 (1026) 
                   
                 (1027), 
                 YNDYXXSX 
                 WFDP 
               
               
                   
                   
                   
                   
                 SQSATWN 
                 KS (1030) 
                 (1024) 
               
               
                   
                   
                   
                   
                 (1028) 
                   
                   
               
               
                   
                   
                   
                   
                 SQSXTWN 
                   
                   
               
               
                   
                   
                   
                   
                 (1029) 
                   
                   
               
               
                   
               
               
                 77D12 
                 SESNXDXX 
                 [none] 
                 XTWDDNX 
                 XFYXH 
                 WINPDXGG 
                 EXXXXXXT 
               
               
                   
                 TNXXN 
                   
                 NXPX 
                 (1037) 
                 TNYAQKFQ 
                 Y (1043) 
               
               
                   
                 (1031), 
                   
                 (1034), 
                   
                 G (1038), 
                   
               
               
                   
                 SESQSDIG 
                   
                 ATWDDNL 
                   
                 WINPESGG 
                   
               
               
                   
                 TNAVN 
                   
                 QGPL 
                   
                 TNYAQKFQ 
                   
               
               
                   
                 (1032) 
                   
                 (1035) 
                   
                 G (1039), 
                   
               
               
                   
                 SESQSDXX 
                   
                 XTWDDNX 
                   
                 WXNPDSX 
                   
               
               
                   
                 TNXXN 
                   
                 QXPX 
                   
                 XTNYXQKF 
                   
               
               
                   
                 (1033) 
                   
                 (1036) 
                   
                 QX (1040), 
                   
               
               
                   
                   
                   
                   
                   
                 WXNPDXX 
                   
               
               
                   
                   
                   
                   
                   
                 XTNYXQKF 
                   
               
               
                   
                   
                   
                   
                   
                 QX (1041), 
                   
               
               
                   
                   
                   
                   
                   
                 WXNPESX 
                   
               
               
                   
                   
                   
                   
                   
                 XTNYXQKF 
                   
               
               
                   
                   
                   
                   
                   
                 QX (1042) 
                   
               
               
                   
               
               
                 87H11 
                 RXSQSXSS 
                 XXSSXQS 
                 [none] 
                 XYYWS 
                 EXNXSXRT 
                 DXGWHFS 
               
               
                   
                 YXN (1044) 
                 (647) 
                   
                 (1045) 
                 NYNPSXKS 
                 FDI (1049), 
               
               
                   
                   
                   
                   
                   
                 (1046), 
                 ESGWHFS 
               
               
                   
                   
                   
                   
                   
                 EIQHSGRT 
                 FDI (1050), 
               
               
                   
                   
                   
                   
                   
                 NYNPSLKS 
                 DSXWHFSF 
               
               
                   
                   
                   
                   
                   
                 (1047) 
                 DX (1051), 
               
               
                   
                   
                   
                   
                   
                 EXQHSXRT 
                 ESXWHFSF 
               
               
                   
                   
                   
                   
                   
                 NYNPSXKS 
                 DX (1052) 
               
               
                   
                   
                   
                   
                   
                 (1048) 
                   
               
               
                   
               
               
                 88H9 
                 RXSQXXRN 
                 XXSXXQX 
                 XQHNSYPT 
                 SYXXH 
                 VIWFDXSN 
                 ERWFXEXX 
               
               
                   
                 DLX (1053) 
                 (1054) 
                 WT (1055), 
                 (639) 
                 EYYADXVK 
                 DY (1063) 
               
               
                   
                   
                   
                 LQHQSYPT 
                   
                 G (1058), 
                   
               
               
                   
                   
                   
                 WT (1056) 
                   
                 VIWFEGSN 
                   
               
               
                   
                   
                   
                 XQHQSYPT 
                   
                 EYYADSVK 
                   
               
               
                   
                   
                   
                 WT (1057) 
                   
                 G (1059), 
                   
               
               
                   
                   
                   
                   
                   
                 VIWFDGSN 
                   
               
               
                   
                   
                   
                   
                   
                 EYYAESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G (1060), 
                   
               
               
                   
                   
                   
                   
                   
                 VIWFEGSN 
                   
               
               
                   
                   
                   
                   
                   
                 EYYAESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G (1061), 
                   
               
               
                   
                   
                   
                   
                   
                 XWFDXSN 
                   
               
               
                   
                   
                   
                   
                   
                 EYYXDSXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (1062) 
                   
               
               
                   
               
               
                 89H9 
                 RXSQSXSY 
                 XXSSXQS 
                 QQSYSSXT 
                 SYXXS 
                 GISGGGGS 
                 EXXXXFDX 
               
               
                   
                 YXN (1064) 
                 (647) 
                 (1065) 
                 (994) 
                 TYYADXVK 
                 (1071) 
               
               
                   
                   
                   
                   
                 G 
                   
                   
               
               
                   
                   
                   
                   
                 (1066), GIS 
                   
                   
               
               
                   
                   
                   
                   
                 GGGGSTY 
                   
                   
               
               
                   
                   
                   
                   
                 YAESVKG 
                   
                   
               
               
                   
                   
                   
                   
                 (1067), 
                   
                   
               
               
                   
                   
                   
                   
                 XXSXXXXS 
                   
                   
               
               
                   
                   
                   
                   
                 TYYXDSXK 
                   
                   
               
               
                   
                   
                   
                   
                 X (1068), 
                   
                   
               
               
                   
                   
                   
                   
                 XXSXXXXS 
                   
                   
               
               
                   
                   
                   
                   
                 TYYXDXXK 
                   
                   
               
               
                   
                   
                   
                   
                 X 
                   
                   
               
               
                   
                   
                   
                   
                 (1069), XXS 
                   
                   
               
               
                   
                   
                   
                   
                 XXXXSTYY 
                   
                   
               
               
                   
                   
                   
                   
                 XESXKX 
                   
                   
               
               
                   
                   
                   
                   
                 (1070) 
                   
                   
               
               
                   
               
               
                 91D7 
                 RXSQNXSS 
                 TXSSXQS 
                 [none] 
                 XYYWS 
                 EXNPXXRT 
                 DNXWHYX 
               
               
                   
                 YXN (1072) 
                 (1073) 
                   
                 (1045) 
                 NYKPSXKS 
                 FDX (1075), 
               
               
                   
                   
                   
                   
                   
                 (1074) 
                 DQGWHYA 
               
               
                   
                   
                   
                   
                   
                   
                 FDI (1076), 
               
               
                   
                   
                   
                   
                   
                   
                 DQXWHYX 
               
               
                   
                   
                   
                   
                   
                   
                 FDX (1077) 
               
               
                   
               
               
                 91F10_LC1 
                 RSSQSXXY 
                 KVSNWDX 
                 XQXTHWP 
                 XYYXH 
                 WXNPNXX 
                 XXSWEXFD 
               
               
                   
                 SDXNTYXN 
                 (1081), KVS 
                 (1086) 
                 (1087) 
                 XTNYXQKF 
                 Y (1091) 
               
               
                   
                 (1078), 
                 NWES 
                   
                   
                 RX (1088), 
                   
               
               
                   
                 RSSQSLVY 
                 (1082), 
                   
                   
                 WINPQSGG 
                   
               
               
                   
                 SDGQTYLN 
                 KXSNWDS 
                   
                   
                 TNYAQKFR 
                   
               
               
                   
                 (1079), 
                 (1083), 
                   
                   
                 G (1089), 
                   
               
               
                   
                 RSSQSXXY 
                 KXSNWDX 
                   
                   
                 WXNPQSX 
                   
               
               
                   
                 SDXQTYXN 
                 (1084), 
                   
                   
                 XTNYXQKF 
                   
               
               
                   
                 (1080) 
                 KXSNWES 
                   
                   
                 RX (1090) 
                   
               
               
                   
                   
                 (1085) 
                   
                   
                   
                   
               
               
                   
               
               
                 91F10_LC2 
                 RXSQSXSS 
                 XXSRXQS 
                 QQSDSTPX 
                 XYYXH 
                 WXNPNXX 
                 XXSWEXFD 
               
               
                   
                 YXN (1044) 
                 (1092) 
                 T (1093) 
                 (1087) 
                 XTNYXQKF 
                 Y (1091) 
               
               
                   
                   
                   
                   
                   
                 RX (1088), 
                   
               
               
                   
                   
                   
                   
                   
                 WINPQSGG 
                   
               
               
                   
                   
                   
                   
                   
                 TNYAQKFR 
                   
               
               
                   
                   
                   
                   
                   
                 G (1089), 
                   
               
               
                   
                   
                   
                   
                   
                 WXNPQSX 
                   
               
               
                   
                   
                   
                   
                   
                 XTNYXQKF 
                   
               
               
                   
                   
                   
                   
                   
                 RX (1090) 
                   
               
               
                   
               
               
                 91G8 
                 RXSQSXSS 
                 XXSSXQS 
                 QQSFSTXT 
                 SYXXS 
                 GISGSGGS 
                 EXXXXFDX 
               
               
                   
                 YXN (1044) 
                 (647) 
                 (1094) 
                 (994) 
                 RYYADXVK 
                 (1071) 
               
               
                   
                   
                   
                   
                   
                 G (1095), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSXSXXS 
                   
               
               
                   
                   
                   
                   
                   
                 RYYXDSXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (1096), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSXSXXS 
                   
               
               
                   
                   
                   
                   
                   
                 RYYXDXXK 
                   
               
               
                   
                   
                   
                   
                   
                 X (1097) 
                   
               
               
                   
               
               
                 92D3 
                 RXSQSXSS 
                 XXSRXQS 
                 QQSDXTPI 
                 XYYXH 
                 WXXPNXX 
                 XSSWEXFD 
               
               
                   
                 YXN (1044) 
                 (1098) 
                 T (1099), 
                 (1087) 
                 XTNYXQKF 
                 Y (1105) 
               
               
                   
                   
                   
                 QQSDTTPX 
                   
                 QX (1102), 
                   
               
               
                   
                   
                   
                 T (1100) 
                   
                 WIIPQSGG 
                   
               
               
                   
                   
                   
                 QQSDXTPX 
                   
                 TNYAQKFQ 
                   
               
               
                   
                   
                   
                 T (1101) 
                   
                 G (1103), 
                   
               
               
                   
                   
                   
                   
                   
                 WXXPQSX 
                   
               
               
                   
                   
                   
                   
                   
                 XTNYXQKF 
                   
               
               
                   
                   
                   
                   
                   
                 QX (1104) 
                   
               
               
                   
               
               
                 92E5 
                 RXSQSXSH 
                 XXSSXQS 
                 QQSFSSXT 
                 SYXXS 
                 GISGRGGS 
                 EXXXXFDX 
               
               
                   
                 YXN (1106) 
                 (647) 
                 (1107) 
                 (994) 
                 TYYADXVK 
                 (1071) 
               
               
                   
                   
                   
                   
                   
                 G (1108), 
                   
               
               
                   
                   
                   
                   
                   
                 GISGRGGS 
                   
               
               
                   
                   
                   
                   
                   
                 TYYAESVK 
                   
               
               
                   
                   
                   
                   
                   
                 G (1109). 
                   
               
               
                   
                   
                   
                   
                   
                 XXSXRXXS 
                   
               
               
                   
                   
                   
                   
                   
                 TYYXDSVK 
                   
               
               
                   
                   
                   
                   
                   
                 X (1110), 
                   
               
               
                   
                   
                   
                   
                   
                 XXSXRXXS 
                   
               
               
                   
                   
                   
                   
                   
                 TYYXDXVK 
                   
               
               
                   
                   
                   
                   
                   
                 X 
                   
               
               
                   
                   
                   
                   
                   
                 (1111), XXS 
                   
               
               
                   
                   
                   
                   
                   
                 XRXXSTYY 
                   
               
               
                   
                   
                   
                   
                   
                 XESVKX 
                   
               
               
                   
                   
                   
                   
                   
                 (1112) 
               
               
                   
               
            
           
         
       
     
     Also preferred are antigen binding proteins comprising the CDR germline sequences related to the antibodies shown in Table A. Such sequences are shown in Table C. 
     
       
         
           
               
             
               
                 TABLE C 
               
             
            
               
                   
               
               
                 Related germline CDR sequences of anti-TL1A antibodies 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Antibody 
                   
                   
                   
                   
                   
                   
               
               
                 designation 
                 LCDR1 
                 LCDR2 
                 LCDR3 
                 HCDR1 
                 HCDR2 
                 HCDR3 
               
               
                 related to 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
               
               
                 germline 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
               
               
                   
               
               
                 3C6 
                 RASQGIRN 
                 AASSLQS 
                 LQHNSYP 
                 SYAMH 
                 VISYDGSN 
                 TTVTYYYY 
               
               
                   
                 DLG (226) 
                 (112) 
                 WT (228) 
                 (253) 
                 KYYADSVK 
                 YGMD (255) 
               
               
                   
                   
                   
                   
                   
                 G (254) 
                   
               
               
                   
               
               
                 2G11 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTP 
                 SYYWS 
                 YIYYSGST 
                 SYYYFD 
               
               
                   
                 YLN (229) 
                 (112) 
                 WT (231) 
                 (256) 
                 NYNPSLKS 
                 (258) 
               
               
                   
                   
                   
                   
                   
                 (172) 
                   
               
               
                   
               
               
                 9C8 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 SYYWS 
                 YIYYSGST 
                 LTGYFD 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (259) 
                 NYNPSLKS 
                 (261) 
               
               
                   
                   
                   
                   
                   
                 (260) 
                   
               
               
                   
               
               
                 23B3 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSTPL 
                 SNSAAWN 
                 RTYYRSK 
                 RDGYNYYY 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 T (237) 
                 (262) 
                 WYNDYAV 
                 YYYGMD 
               
               
                   
                 A (235) 
                   
                   
                   
                 SVKS (184) 
                 (264) 
               
               
                   
               
               
                 23B3 VH4 
                 RASQGISN 
                 AASTLQS 
                 QKYNSAPL 
                 SGSYYWS 
                 YIYYSGST 
                 RDGYNYYY 
               
               
                   
                 YLA (241) 
                 (242) 
                 T (243) 
                 (268) 
                 NYNPSLKS 
                 YYYGMD 
               
               
                   
                   
                   
                   
                   
                 (269) 
                 (270) 
               
               
                   
               
               
                 23B3 VH3 
                 RASQGISN 
                 AASTLQS 
                 QKYNSAPL 
                 SNYMS 
                 IYSGGSTY 
                 RDGYNYYY 
               
               
                   
                 YLA (241) 
                 (242) 
                 T (243) 
                 (271) 
                 YADSVKG 
                 YYYGMD 
               
               
                   
                   
                   
                   
                 (272) 
                 (273) 
                   
               
               
                   
               
               
                 3B3 
                 RASQSVSS 
                 GASSRAT 
                 QQYGSSPI 
                 GYYWS 
                 EINHSGST 
                 GYCSSTSC 
               
               
                   
                 SYLA (238) 
                 (124) 
                 T (240) 
                 (265) 
                 NYNPSLKS 
                 YTYFD 
               
               
                   
                   
                   
                   
                   
                 (266) 
                 (267) 
               
               
                   
               
               
                 5G4 
                 RASQSVSS 
                 GASSRAT 
                 QQYGSSPI 
                 GYYWS 
                 EINHSGST 
                 GYCSSTSC 
               
               
                   
                 SYLA (238) 
                 (124) 
                 T (240) 
                 (265) 
                 NYNPSLKS 
                 YTYFDY 
               
               
                   
                   
                   
                   
                   
                 (266) 
                 (672) 
               
               
                   
               
               
                 17E9 
                 RASQGIRN 
                 AASSLQS 
                 LQHNSYPL 
                 SYAMH 
                 VISYDGSN 
                 GTTGTYYY 
               
               
                   
                 DLG (226) 
                 (112) 
                 T (674) 
                 (253) 
                 KYYADSVK 
                 YYGMDV 
               
               
                   
                   
                   
                   
                   
                 G (254) 
                 (675) 
               
               
                   
               
               
                 53D3 
                 RSSQSLLH 
                 LGSNRAS 
                 MQALQTPL 
                 SYSMN 
                 SISSSSSYI 
                 GIAAAGYY 
               
               
                   
                 SNGYNYLD 
                 (935) 
                 T (936) 
                 (950) 
                 YYADSVKG 
                 YYYGMDV 
               
               
                   
                 (1113) 
                   
                   
                   
                 (951) 
                 (952) 
               
               
                   
               
               
                 54E5 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSTP 
                 SYGMH 
                 VIWYDGSN 
                 EYSSSSYY 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 WT (938) 
                 (164) 
                 KYYADSVK 
                 YYYGMDV 
               
               
                   
                 A (235) 
                   
                   
                   
                 G (953) 
                 (954) 
               
               
                   
               
               
                 56E1 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSTP 
                 SYGMH 
                 VIWYDGSN 
                 EYSSSSYY 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 WT (938) 
                 (164) 
                 KYYADSVK 
                 YYYGMDV 
               
               
                   
                 A (235) 
                   
                   
                   
                 G (953) 
                 (954) 
               
               
                   
               
               
                 57A8 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 GTTGTYYY 
               
               
                   
                 AGYDVH 
                 (148) 
                 GWV (701) 
                 (836) 
                 TYYADSVK 
                 YYGMDV 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                 (675) 
               
               
                   
               
               
                 58G5 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 RDGYNYYF 
               
               
                   
                 AGYDVH 
                 (148) 
                 GYV (707) 
                 (836) 
                 TYYADSVK 
                 DY (958) 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 60G11 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 RDGYNYYF 
               
               
                   
                 AGYDVH 
                 (148) 
                 GYV (707) 
                 (836) 
                 TYYADSVK 
                 DY (958) 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 73C2 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 SNSAAWN 
                 RTYYRSK 
                 GIAVAGNW 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (262) 
                 WYNDYAV 
                 FDP (961) 
               
               
                   
                   
                   
                   
                   
                 SVKS (184) 
                   
               
               
                   
               
               
                 76A4 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 SNSAAWN 
                 RTYYRSK 
                 GIAVAGNW 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (262) 
                 WYNDYAV 
                 FDP (961) 
               
               
                   
                   
                   
                   
                   
                 SVKS (184) 
                   
               
               
                   
               
               
                 77D12 
                 SGSSSNIG 
                 SNNQRPS 
                 AAWDDSL 
                 GYYMH 
                 WINPNSGG 
                 SIAARAEYF 
               
               
                   
                 SNTVN 
                 (948) 
                 NGVV 
                 (857) 
                 TNYAQKFQ 
                 QH (964) 
               
               
                   
                 (947) 
                   
                 (949) 
                   
                 G (963) 
                   
               
               
                   
               
               
                 87H11 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 GYYWS 
                 EINHSGST 
                 DYGDYDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (265) 
                 NYNPSLKS 
                 DI (966) 
               
               
                   
                   
                   
                   
                   
                 (266) 
                   
               
               
                   
               
               
                 88H9 
                 RASQGIRN 
                 AASSLQS 
                 LQHNSYP 
                 SYGMH 
                 VIWYDGSN 
                 RWLQLYFD 
               
               
                   
                 DLG (226) 
                 (112) 
                 WT (228) 
                 (164) 
                 KYYADSVK 
                 Y (967) 
               
               
                   
                   
                   
                   
                   
                 G (953) 
                   
               
               
                   
               
               
                 89H9 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 VEMATIDA 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 FDI (968) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 91D7 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 GYYWS 
                 EINHSGST 
                 DYGDYDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (265) 
                 NYNPSLKS 
                 DI (966) 
               
               
                   
                   
                   
                   
                   
                 (266) 
                   
               
               
                   
               
               
                 91F10_LC1 
                 RSSQSLVY 
                 KVSNRDS 
                 MQGTHWP 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 SDGNTYLN 
                 (943) 
                 LT (944) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                 (753) 
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 91F10_LC2 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                   
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 91G8 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 SIAARDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 DI (970) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 92D3 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                   
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 92E5 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 SIAARDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 DI (970) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 88H9 
                 RASQGIRN 
                 AASSLQS 
                 LQHNSYP 
                 SYGMH 
                 VIWYDGSN 
                 RWLQLYFD 
               
               
                   
                 DLG (226) 
                 (112) 
                 WT (228) 
                 (164) 
                 KYYADSVK 
                 Y (967) 
               
               
                   
                   
                   
                   
                   
                 G (953) 
                   
               
               
                   
               
               
                 54E5 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSTP 
                 SYGMH 
                 VIWYDGSN 
                 EYSSSSYY 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 WT (938) 
                 (164) 
                 KYYADSVK 
                 YYYGMDV 
               
               
                   
                 A (235) 
                   
                   
                   
                 G (953) 
                 (954) 
               
               
                   
               
               
                 56E1 
                 KSSQSVLY 
                 WASTRES 
                 QQYYSTP 
                 SYGMH 
                 VIWYDGSN 
                 EYSSSSYY 
               
               
                   
                 SSNNKNYL 
                 (118) 
                 WT (938) 
                 (164) 
                 KYYADSVK 
                 YYYGMDV 
               
               
                   
                 A (235) 
                   
                   
                   
                 G (953) 
                 (954) 
               
               
                   
               
               
                 53D3 
                 RSSQSLLH 
                 LGSNRAS 
                 MQALQTPL 
                 SYSMN 
                 SISSSSSYI 
                 GIAAAGYY 
               
               
                   
                 SNGYNYLD 
                 (935) 
                 T (936) 
                 (950) 
                 YYADSVKG 
                 YYYGMDV 
               
               
                   
                 (1113) 
                   
                   
                   
                 (951) 
                 (952) 
               
               
                   
               
               
                 89H9 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 VEMATIDA 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 FDI (968) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 91G8 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 SIAARDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 DI (970) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 92E5 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 SYAMS 
                 AISGSGGS 
                 SIAARDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (836) 
                 TYYADSVK 
                 DI (970) 
               
               
                   
                   
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 92D3 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                   
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 91F10_LC2 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPI 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (942) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                   
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 91D7 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 GYYVVS 
                 EINHSGST 
                 DYGDYDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (265) 
                 NYNPSLKS 
                 DI (966) 
               
               
                   
                   
                   
                   
                   
                 (266) 
                   
               
               
                   
               
               
                 76A4 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 SNSAAWN 
                 RTYYRSK 
                 GIAVAGNW 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (262) 
                 WYNDYAV 
                 FDP (961) 
               
               
                   
                   
                   
                   
                   
                 SVKS (184) 
                   
               
               
                   
               
               
                 87H11 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 GYYWS 
                 EINHSGST 
                 DYGDYDAF 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (265) 
                 NYNPSLKS 
                 DI (966) 
               
               
                   
                   
                   
                   
                   
                 (266) 
                   
               
               
                   
               
               
                 73D2 
                 RASQSISS 
                 AASSLQS 
                 QQSYSTPY 
                 SNSAAWN 
                 RTYYRSK 
                 GIAVAGNW 
               
               
                   
                 YLN (229) 
                 (112) 
                 T (234) 
                 (262) 
                 WYNDYAV 
                 FDP (961) 
               
               
                   
                   
                   
                   
                   
                 SVKS (184) 
                   
               
               
                   
               
               
                 91F10_LC1 
                 RSSQSLVY 
                 KVSNRDS 
                 MQGTHWP 
                 GYYMH 
                 WINPNSGG 
                 GYSYGYYF 
               
               
                   
                 SDGNTYLN 
                 (943) 
                 LT (944) 
                 (857) 
                 TNYAQKFQ 
                 DY (969) 
               
               
                   
                 (753) 
                   
                   
                   
                 G (963) 
                   
               
               
                   
               
               
                 60G11 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 RDGYNYYF 
               
               
                   
                 AGYDVH 
                 (148) 
                 GYV (707) 
                 (836) 
                 TYYADSVK 
                 DY (958) 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 58G5 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 RDGYNYYF 
               
               
                   
                 AGYDVH 
                 (148) 
                 GYV (707) 
                 (836) 
                 TYYADSVK 
                 DY (958) 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                   
               
               
                   
               
               
                 57A8 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYAMS 
                 AISGSGGS 
                 GTTGTYYY 
               
               
                   
                 AGYDVH 
                 (148) 
                 GWV (701) 
                 (836) 
                 TYYADSVK 
                 YYGMDV 
               
               
                   
                 (146) 
                   
                   
                   
                 G (956) 
                 (675) 
               
               
                   
               
               
                 77D12 
                 SGSSSNIG 
                 SNNQRPS 
                 AAWDDSL 
                 GYYMH 
                 WINPNSGG 
                 SIAARAEYF 
               
               
                   
                 SNTVN 
                 (948) 
                 NGVV (142) 
                 (857) 
                 TNYAQKFQ 
                 QH (964) 
               
               
                   
                 (947) 
                   
                   
                   
                 G (963) 
               
               
                   
               
            
           
         
       
     
     Further preferred are antigen binding proteins comprising the variable domain sequences of the preferred anti-TL1A antibodies, as shown in Table D. Throughout, “VH” as shown in Table D refers to the variable domain of the heavy chain, “VL” to that of the light chain. Molecules within this invention may incorporate modifications to the sequences shown in Table D, including truncations or substitutions for stability or other functionality or removal of hotspots (chemical or physical modification of amino acids). Also included within the invention are molecules having at least 90% sequence identity with the sequences shown in Table D. 
     
       
         
           
               
             
               
                 TABLE D 
               
             
            
               
                   
               
               
                 Variable domain sequences of preferred anti-TL1A antibodies 
               
               
                 Antibody 
               
            
           
           
               
               
            
               
                 designation 
                 Amino acid sequence (SEQ ID NO) 
               
               
                   
               
               
                 3C6 VL 
                 DIQMTQSPSSLSASVGDRVTITCRTSQDIRDDLGWYQQKPGKAPKRLIYDASSLQSGV 
               
               
                   
                 PSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPPTFGQGTKVEIK (6) 
               
               
                   
               
               
                 3C6 VH 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDG 
               
               
                   
                 NNKLYTDSVKGRFTISRDDSKSTLYLQMNSLRAEDTAVYYCARDPTVTLYYYYGMDV 
               
               
                   
                 WGQGTTVTVSS (8) 
               
               
                   
               
               
                 2G11 VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSINNYLNWYQQKPGIAPKLLIYATSSLQSGVP 
               
               
                   
                 SRFSGSGSGTDFSLTISSLQPEDFATYFCQQSYSTPRTFGQGTKVEIK (10) 
               
               
                   
               
               
                 2G11 VH 
                 QVQLQESGPGLVKPSETLSLTCTVSGGSISSYFWSWIRQPPGKGLEWIGYIYYSGSTN 
               
               
                   
                 YNPSLKSRVTMSIDTSKNQFSLKLSSVTAADTAVYYCAREIGSYYGFDYWGQGALVTV 
               
               
                   
                 SS (12) 
               
               
                   
               
               
                 9C8 VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSINNYLNWYQQRPGKAPKLLIYAASSLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTISSLQPGDFATYYCQQSYSTPRTFGQGTKLEIK (14) 
               
               
                   
               
               
                 9C8 VH 
                 QVQLQESGPGLVKPSETLSLTCTVSGGSISSYFWSWIRQPPGKGLEWIGYIYYSGNTK 
               
               
                   
                 YNPSLKSRVTISIDTSKNQFSLKLSSVTAADTAVYYCARETGSYYGFDYWGQGTLVTV 
               
               
                   
                 SS (16) 
               
               
                   
               
               
                 23B3 VL 
                 DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLVWYQQKPGQPPKLLIYWAS 
               
               
                   
                 TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYKTPLTFGGGTKVEIK (18) 
               
               
                   
               
               
                 23B3 VH 
                 QVQLQQSGPGLVKPSQTLSLTCVISGDSVSTNSVAWNWIRQSPSRGLEWLGRTYYR 
               
               
                   
                 SKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREDGDSYYRYGMD 
               
               
                   
                 VWGQGTTVTVSS (20) 
               
               
                   
               
               
                 3B3 VL 
                 EIVLTQSPGTLSLSPGERATLSCRASQSVRSSYLAWYQQKPGQAPRLLIYGASSRATG 
               
               
                   
                 IPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPTFGQGTRLEIK (22) 
               
               
                   
               
               
                 3B3 VH 
                 QVQLQQWGAGLLKPSETLSLTCAVHGGSFSGYYWNWIRQPPGKGLEWIGEINHAGN 
               
               
                   
                 TNYNPSLKSRVTISLDTSKNQFSLTLTSVTAADTAVYYCARGYCRSTTCYFDYWGQGT 
               
               
                   
                 LVTVSS (24) 
               
               
                   
               
               
                 23B3 VH4 
                 DIQMTQSPSSLSASVGDRVTITCRSSQSVLYSSNNKNYLVWYQQKPGKVPKLLIYWAS 
               
               
                 VL 
                 TRESGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQYYKTPLTFGGGTKVEIK (26) 
               
               
                   
               
               
                 23B3 VH4 
                 QVQLQESGPGLVKPSETLSLTCTISGDSVSTNSVAWNWIRQPPGKGLEWIGRTYYRS 
               
               
                 VH 
                 KWYNDYAVSLKSRVTISPDTSKNQFSLKLSSVTAADTAVYYCAREDGDSYYRYGMDV 
               
               
                   
                 WGQGTTVTVSS (28) 
               
               
                   
               
               
                 23B3 VH3 
                 DIQMTQSPSSLSASVGDRVTITCRSSQSVLYSSNNKNYLVWYQQKPGKVPKLLIYWAS 
               
               
                 VL 
                 TRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYKTPLTFGGGTKVEIK (30) 
               
               
                   
               
               
                 23B3 VH3 
                 EVQLLESGGGLVQPGGSLRLSCAISGDSVSTNSVAWNWIRQAPGKGLEWVSRTYYR 
               
               
                 VH 
                 SKWYNDYAVSVKGRFTISPDTSKNIFYLQMNSLRAEDTAVYYCAREDGDSYYRYGM 
               
               
                   
                 DVWGQGTTVTVSS (32) 
               
               
                   
               
               
                 5G4 VL 
                 EIVLTQSPGTLSLSPGERVTLSCRASQSVRSSYLAWYQQRAGQAPRLLIYGASSRATG 
               
               
                   
                 IPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPTFGQGTRLEIK (491) 
               
               
                   
               
               
                 5G4 VH 
                 QVQLQQWGAGLLKPSETLSLTCDVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGIT 
               
               
                   
                 NYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGYCRSTTCYFDYWGQGT 
               
               
                   
                 LVTVSS (495) 
               
               
                   
               
               
                 17E9 VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQGISNDLAWYQQKPGKAPKRLIFAASSLQSGV 
               
               
                   
                 PSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPPTFGGGTKVEIK (493) 
               
               
                   
               
               
                 17E9 VH 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTLSSYGMHWVRQAPGKGLEWVAVMSYDG 
               
               
                   
                 NNKLYADSVKGRFTISRDNSKKTLYLQMNSLRAEDTAVYYCARDETETLYYYYGIDVW 
               
               
                   
                 GQGTTVTVSS (497) 
               
               
                   
               
               
                 53D3 VL 
                 DIVMTQSPLSLPVTPGEPASISCRSSQSLLYSNGYNYLDWYLQKPGQSPQLLIYLGSS 
               
               
                   
                 RASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQPLQTPLTFGGGTKVEIK (866) 
               
               
                   
               
               
                 53D3 VH 
                 EVQLVESGGGLVKPGGSLRLSCAASEFTFSTYYMSWVRQAPGKGLEWVSSISSSSSF 
               
               
                   
                 IYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDRIAAPGTYYYYGMDVW 
               
               
                   
                 GQGTTVTVSS (868) 
               
               
                   
               
               
                 54E5 VL 
                 DIVMTQSPDSLAVSLGERATINCKSSQNILYSSNNKNYLAWYQQKPGQPPSLLIYWAS 
               
               
                   
                 TRESGVPDRFSGSGSGTDFTLTLSSLQAEDVAVYFCQQYYSIPWTFGQGTKVEIK 
               
               
                   
                 (870) 
               
               
                   
               
               
                 54E5 VH 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWAVIWYDG 
               
               
                   
                 SNKDYADSVKGRFTVSRDNSRDTLYLQMNSLRAEDTAVYYCAREERDSYYHYGMDV 
               
               
                   
                 WGQGTTVTVSS (872) 
               
               
                   
               
               
                 56E1 VL 
                 DIVMTQSPDSLTVSLGEGATINCKSSQSVLYSSNNKNYLAWYQQKPGQPPSLLIYWAS 
               
               
                   
                 TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSIPWTFGQGTKVEIK 
               
               
                   
                 (874) 
               
               
                   
               
               
                 56E1 VH 
                 QVQLVESGGGVVQPGRSLRLSCAASGFIFSSYGMHWVRQAPGKGLEWVAVIWYDGS 
               
               
                   
                 NKDYADSVKGRFTVSRDNSRDTLYLQMNSLRAEDTAVYYCAREERDSYYHYGMDVW 
               
               
                   
                 GQGTTVTVSS (876) 
               
               
                   
               
               
                 57A8VL 
                 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYNVHWYQQLPGTAPKLLIYGNNNRP 
               
               
                   
                 SGVPDRFSGSKSGTSASLAITGLQTEDEADYYCQSYDSSLSGWVFGGGTKLTVL 
               
               
                   
                 (878) 
               
               
                   
               
               
                 57A8VH 
                 EIQLLESGGGLVQPGGSLRLSCVASGFTFSSYVMSWVRQAPGKGLEWVSVISGSGG 
               
               
                   
                 STYYADSVKGRFTISRDNSKTTLYLQMNSLRAEDTAVYYCAKGGTNYYYYYSGMDVW 
               
               
                   
                 GQGTTVTVSS (880) 
               
               
                   
               
               
                 58G5VL 
                 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSHRP 
               
               
                   
                 SGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVL (882) 
               
               
                   
               
               
                 58G5VH 
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLQWVSVISGRGG 
               
               
                   
                 STYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDGYSSAWYFDYWGQ 
               
               
                   
                 GTLVTVSS (884) 
               
               
                   
               
               
                 60G11VL 
                 QSVLTQPPSVSGAPGQRVTISCTGSGSNIGAGYDVHWYQQLPGTAPKWYGNSHRP 
               
               
                   
                 SGIPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVL (886) 
               
               
                   
               
               
                 60G11VH 
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAMNWVRQAPGKGLEWVSVISGRGG 
               
               
                   
                 STYYADSVKGRFTISRDNSRNTLYLQMNSLRAEDTAIYYCAKDGYSSAWFFDYWGQG 
               
               
                   
                 TLITVSS (888) 
               
               
                   
               
               
                 73C2VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSFSSYLNWYQQKPGKAPELLIYAASSLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTISSLQPEDFASYFCQQSYSTPRSFGQGTKLEIK (890) 
               
               
                   
               
               
                 7302VH 
                 QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSSSATWNWIRQSPSRGLEWLGRTYQR 
               
               
                   
                 SKWNNDYAVSVKSRITINPDTSRNQFSLQLNSVTPEDTAVYYCAREVVAGPRWFDPW 
               
               
                   
                 GQGTLVTVSS (892) 
               
               
                   
               
               
                 76A4VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSVISYLNWYQQKPGKAPKLLIYTASSLQSGIP 
               
               
                   
                 SRFSGSGSGTDFTLTISSLQPEDFATYFCQQSYSTPRSFGQGTKLEIK (894) 
               
               
                   
               
               
                 76A4VH 
                 QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYR 
               
               
                   
                 SKLYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVVAGPRWFDPW 
               
               
                   
                 GQGTLVTVSS (896) 
               
               
                   
               
               
                 77D12VL 
                 QSVLTQPPSASGTPGQRVTISCSESNSDIGTNAVNWYQQFPGTAPKFLIYSNNKRPSG 
               
               
                   
                 VPDRFSGSKSGTSASLAISGLQSEDEADYYCATWDDNLNGPLFGGGTKLTVL (898) 
               
               
                   
               
               
                 77D12VH 
                 QVQLVQSGAEVKKPGASVRVSCKASGYTFTGFYMHWVRQAPGQGLEWMGWINPDS 
               
               
                   
                 GGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCAREGIAVALTYWGQGT 
               
               
                   
                 LVTVSS (900) 
               
               
                   
               
               
                 87H11VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYVASSLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSNPQECSFGQGTKLEIK (902) 
               
               
                   
               
               
                   
               
               
                 87H11VH 
                 QEQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGR 
               
               
                   
                 TNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARDSGWHFSFDIWDQGTM 
               
               
                   
                 VTVSS (904) 
               
               
                   
               
               
                 88H9VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQEKPGKAPKRLIYAASGLQGG 
               
               
                   
                 DPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPTWTFGQGTKVEIK (906) 
               
               
                   
               
               
                 88H9VH 
                 QVQLVESGAGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGMGLEWVAVIWFDG 
               
               
                   
                 SNEYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERWFGELLDYWGQ 
               
               
                   
                 GTLVTVSS (908) 
               
               
                   
               
               
                 89H9VL 
                 DIQMTQSPSSLSASVGDSVTITCRASQSISYYLNWYQQKPGKAPKFLIYAASSLQSGVP 
               
               
                   
                 SRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSSITFGQGTRLEIK (910) 
               
               
                   
               
               
                 89H9VH 
                 EVHLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSGISGGGG 
               
               
                   
                 STYYADSVKGRFTISRDNSKNTLYLQMNSLRAENTAVYYCAIEMAGAFDIWGQGTMVT 
               
               
                   
                 VSS (912) 
               
               
                   
               
               
                 91D7VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQNISSYLNWYQQRPGKAPNLLLFTASSLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTINSLQPEDFATYYCQQSYSNPPESSFGQGTKLEIK (914) 
               
               
                   
               
               
                 91D7VH 
                 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINPVGR 
               
               
                   
                 TNYKPSLKSRVTISVDTSKNQFSLKLSSVTAADTALYYCARDNGWHYAFDIWGQGTM 
               
               
                   
                 VTVSS (916) 
               
               
                   
               
               
                 91F10_LC1VL 
                 DVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGNTYLNWFQQRPGRSPRRLIYKVS 
               
               
                   
                 NWDSGVPDRFSGSGSGTDSTLIISRVEAEDVGVYYYMQGTHWPLGGGTKVEIK (918) 
               
               
                   
               
               
                 91F10_LC1VH 
                 QGQLVQSGAEVKKPGASVKVSCQASGYTFTAYYMHWVRQAPGQGLEWMGWINPNS 
               
               
                   
                 GGTNYAQKFRGRVTMTRDTSISTAYMELSRLRSDDTAVYYCATGGSWEGFDYWGQ 
               
               
                   
                 GTLVTVSS (920) 
               
               
                   
               
               
                 91F10_LC2VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKFLIYAASRLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSDSTPITFGQGTRLEIK (922) 
               
               
                   
               
               
                 91F10_LC2VH 
                 QGQLVQSGAEVKKPGASVKVSCQASGYTFTAYYMHWVRQAPGQGLEWMGWINPNS 
               
               
                   
                 GGTNYAQKFRGRVTMTRDTSISTAYMELSRLRSDDTAVYYCATGGSWEGFDYWGQ 
               
               
                   
                 GTLVTVSS (920) 
               
               
                   
               
               
                 91G8VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKFLIYAASSLQSGVP 
               
               
                   
                 SRFSGSGSGTDFTLTISSLQPEDFATYYCQQSFSTITFGQGTRLEIK (924) 
               
               
                   
               
               
                 91G8VH 
                 EVQLLESGGGLVQPGGSLRLSCVASGFTFSSYAMSWVRQAPGKGLEWVSGISGSGG 
               
               
                   
                 SRYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAIEVAGAFDIWGQGTMVT 
               
               
                   
                 VSS (926) 
               
               
                   
               
               
                 92D3VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGTAPKFLIYGASRLQSGV 
               
               
                   
                 PSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSDTTPITFGQGTRLEIK (928) 
               
               
                   
               
               
                 9203VH 
                 QVQLVQSGAEVKKPGASVQVSCKASGYTFTGYYMHWIRQAPGQGLEWLGWIIPNSG 
               
               
                   
                 GTNYAQKFQGRVIMTRDTSISTAYMELSRLRSDDTAVYYCATGSSWEGFDYWGQGT 
               
               
                   
                 LVTVSS (930) 
               
               
                   
               
               
                 92E5VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQSISHYLNWYQQKPGKAPKFLIYAASSLQSGVP 
               
               
                   
                 SRFSGSGSGTDFTLTISSLQPEDFATYYCQQSFSSITFGQGTRLEIK (932) 
               
               
                   
               
               
                 92E5VH 
                 EVHLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQVPGKGLEWVSGISGRGG 
               
               
                   
                 STYYADSVKGRFTISRDNSKNTLYLQMDSLRAEDTAVYNCAIEVAGAFDIWGQGTMVT 
               
               
                   
                 VSS (934) 
               
               
                   
               
            
           
         
       
     
     Each of the foregoing polypeptides is encoded by a nucleic acid having the nucleotide sequence immediately preceding the polypeptide sequence in the Sequence Listing. The sequences from antibodies 9C8 and 3B3 are preferred. 
     The full anti-TL1A antibody sequences shown in Table E are preferred for the anti-TL1A antibodies. LC refers to the antibody light chain, HC to the heavy chain. Also encompassed within the invention are molecules having at least 90% sequence identity with either or both of the light chain and heavy chain sequences in Table E. 
     
       
         
           
               
             
               
                 TABLE E 
               
             
            
               
                   
               
               
                 Preferred anti-TL1A Antibodies 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Antibody 
                 Light chain 
                 Heavy chain 
                 iPS (molecule 
               
               
                   
                 designation 
                 SEQ ID NO 
                 SEQ ID NO 
                 designation) 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 3C6 
                 50 
                 52 
                 284112 
               
               
                   
                 2G11 
                 54 
                 56 
                 285396 
               
               
                   
                 9C8 
                 58 
                 60 
                 285412 
               
               
                   
                 23B3 
                 62 
                 64 
                 290043 
               
               
                   
                 3B3 
                 66 
                 68 
                 308844 
               
               
                   
                 23B3 HC4 
                 70 
                 72 
                 325246 
               
               
                   
                 23B3 HC3 
                 74 
                 76 
                 325336 
               
               
                   
                 5G4 
                 455 
                 457 
                 284510 
               
               
                   
                 17E9 
                 459 
                 461 
                 284078 
               
               
                   
                 88H9 
                 1116 
                 1118 
                 427485 
               
               
                   
                 54E5 
                 1120 
                 1122 
                 427576 
               
               
                   
                 60G11 
                 1124 
                 1126 
                 427493 
               
               
                   
                 53D3 
                 1128 
                 1130 
                 427504 
               
               
                   
                 89H9 
                 1132 
                 1134 
                 427509 
               
               
                   
                 91D7 
                 1136 
                 1138 
                 427514 
               
               
                   
                 57A8 
                 1140 
                 1142 
                 427519 
               
               
                   
                 91G8 
                 1144 
                 1146 
                 427524 
               
               
                   
                 76A4 
                 1148 
                 1150 
                 427529 
               
               
                   
                 87H11 
                 1152 
                 1154 
                 427534 
               
               
                   
                 58G5 
                 1156 
                 1158 
                 427539 
               
               
                   
                 57A8 
                 1160 
                 1162 
                 427519 
               
               
                   
                 73C2 
                 1164 
                 1166 
                 427544 
               
               
                   
                 77D12 
                 1168 
                 1170 
                 427549 
               
               
                   
                 56E1 
                 1172 
                 1174 
                 427554 
               
               
                   
                 92E5 
                 1176 
                 1178 
                 427559 
               
               
                   
                 92D3 
                 1180 
                 1182 
                 427564 
               
               
                   
                 91F10_LC1 
                 1184 
                 1186 
                 427580 
               
               
                   
                 91F10_LC2 
                 1188 
                 1190 
                 427572 
               
               
                   
                   
               
            
           
         
       
     
     Each of the amino acid sequences in Table E is encoded by the nucleic acid sequence immediately preceding it in the Sequence Listing. The sequences from antibodies 9C8 and 3B3 are preferred. 
     The foregoing anti-TL1A antibody sequences can be used in bispecific antigen proteins in any of the formats described herein. Preferably, such sequences are used in hetero Ig antigen binding proteins and IgG-scFv bispecific antigen binding proteins as described in this specification. 
     Hetero Ig Bispecific Antigen Binding Proteins 
     The invention further relates to hetero Ig bispecific antigen binding proteins as shown in  FIG. 1  that comprise TL1A-specific binding entities. Structurally, hetero Ig bispecific antigen binding proteins comprise: 
     (a) a TL1A binding entity light chain variable domain comprised in a light chain separate from a TL1A binding entity heavy chain variable domain, a heavy chain variable domain directed to a different antigen (e.g., TNF-α), and a binding entity light chain variable domain directed to the different antigen; 
     (b) a TL1A binding entity heavy chain variable domain comprised in a heavy chain separate from the TL1A binding entity light chain variable domain, the heavy chain variable domain directed to a different antigen, and the light chain variable domain directed to a different antigen; 
     (c) the heavy chain variable domain directed to a different antigen comprised in a heavy chain separate from the light chain domain directed to said different antigen, the TL1A binding entity heavy chain variable domain, and the TL1A binding entity light chain variable domain; 
     (d) the heavy chain comprising the TL1A binding entity heavy chain variable domain covalently bound to the light chain comprising the TL1A binding entity light chain variable domain; 
     (e) the heavy chain comprising the heavy chain variable domain directed to said different antigen covalently bound to the light chain comprising light chain variable domain directed to the different antigen; and 
     (f) the heavy chain comprising the TL1A binding entity heavy chain variable domain covalently bound to the heavy chain comprising the heavy chain variable domain directed to the different antigen. 
     For such hetero Ig bispecific antibodies, the TL1A-specific binding entities are preferred to comprise one or more of the CDRs listed in Table A. The variable region domains in Table D may be employed but it is preferred that the TL1A binding entity comprise charged amino acids that aid in correct assembly of the hetero Ig bispecific antigen binding protein (see  FIG. 1 ). Variable region sequences preferred for the hetero Ig bispecific antibodies are shown in Table F. As before, VL in Table F refers to the light chain variable region, VH to the heavy chain variable region. The SEQ ID NO from the Sequence Listing appears after each sequence in Table F. 
     
       
         
           
               
             
               
                 TABLE F 
               
             
            
               
                   
               
               
                 Hetero Ig TL1A-specific variable regions 
               
            
           
           
               
               
            
               
                 Antibody 
                   
               
               
                 designation 
                 Amino acid sequence (SEQ ID NO) 
               
               
                   
               
               
                 3B3 variant 
                 EIVLTQSPGTLSLSPGERATLSCRASQSVRSSYLAWYQQKPGQAPRLLIYGASSRATG 
               
               
                 VL 
                 IPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPTFGQGTRLEIK (288) 
               
               
                   
               
               
                 3B3 variant 
                 QVQLQQWGAGLLKPSETLSLTCAVHGGSFSGYYWNWIRQPPGKGLEWIGEINHAGIT 
               
               
                 VH 
                 NYNPSLKSRVTISLDTSKNQFSLKLTSVTAADTAVYYCARGYCRSTTCYFDYWGQGTL 
               
               
                   
                 VTVSS (290) 
               
               
                   
               
               
                 2G11 variant 
                 DIQMTQSPSSLSASVGDRVTITCRASQSINNYLNWYQQKPGKAPKLLIYATSSLQSGV 
               
               
                 VL 
                 PSRFSGSGSGTDFSLTISSLQPEDFATYFCQQSYSTPRTFGQGTKVEIK (292) 
               
               
                   
               
               
                 2G11 variant 
                 QVQLQESGPGLVKPSETLSLTCTVSGGSISSYFWSWIRQPPGKGLEWIGYIYYSGSTN 
               
               
                 VH 
                 YNPSLKSRVTMSIDTSKNQFSLKLSSVTAADTAVYYCAREIGSYYGFDYWGQGTLVTV 
               
               
                   
                 SS (294) 
               
               
                   
               
               
                 23B3 VH4 
                 DIQMTQSPSSLSASVGDRVTITCRSSQSVLYSSNNKNYLVWYQQKPGKVPKLLIYWAS 
               
               
                 VL 
                 TRESGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQYYKTPLTFGGGTKVEIK 
               
               
                   
                 (296) 
               
               
                   
               
               
                 23B3 VH4 
                 QVQLQESGPGLVKPSETLSLTCTISGDSVSTNSVAWNWIRQPPGKGLEWIGRTYYRS 
               
               
                 VH 
                 KWYNDYAVSLKSRVTISPDTSKNQFSLKLSSVTAADTAVYYCAREDGDSYYRYGMDV 
               
               
                   
                 WGQGTTVTVSS (298) 
               
               
                   
               
               
                 23B3 VH3 
                 DIQMTQSPSSLSASVGDRVTITCRSSQSVLYSSNNKNYLVWYQQKPGKVPKLLIYWAS 
               
               
                 VL 
                 TRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYKTPLTFGGGTKVEIK 
               
               
                   
                 (300) 
               
               
                   
               
               
                 23B3 VH3 
                 EVQLLESGGGLVQPGGSLRLSCAISGDSVSTNSVAWNWIRQAPGKGLEWVSRTYYR 
               
               
                 VH 
                 SKWYNDYAVSVKGRFTISPDTSKNIFYLQMNSLRAEDTAVYYCAREDGDSYYRYGM 
               
               
                   
                 DVWGQGTTVTVSS (302) 
               
               
                   
               
               
                 3C6 variant 
                 DIQMTQSPSSLSASVGDRVTITCRTSQDIRDDLGWYQQKPGKAPKLLIYDASSLQSGV 
               
               
                 VL 
                 PSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPPTFGQGTKVEIK (304) 
               
               
                   
               
               
                 3C6 variant 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDG 
               
               
                 VH 
                 NNKLYTDSVKGRFTISRDDSKSTLYLQMNSLRAEDTAVYYCARDPTVTLYYYYGMDV 
               
               
                   
                 WGQGTTVTVSS (306) 
               
               
                   
               
               
                 3C6 
                 DIQMTQSPSSLSASVGDRVTITCRTSQDIRDDLGWYQQKPGKAPKRLIYDASSLQSGV 
               
               
                 VL 
                 PSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPPTFGQGTKVEIK (308) 
               
               
                   
               
               
                 3C6 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDG 
               
               
                 VH 
                 NNKLYTDSVKGRFTISRDDSKSTLYLQMNSLRAEDTAVYYCARDPTVTLYYYYGMDV 
               
               
                   
                 WGQGTTVTVSS (310) 
               
               
                   
               
            
           
         
       
     
     Each of the polypeptides in Table F is encoded by a nucleic acid having the sequence immediately preceding the polypeptide sequence in the Sequence Listing. 
     Within the scope of this invention are hetero IgG molecules with a TNF-specific binding entity, preferably together with a TL1A-specific binding entity. The TNF-specific binding entity preferably comprises one or more sequences of CDRs or variable domains of antibodies 3.2 (described hereinafter), 234 (described hereinafter), certolizumab, adalimumab, infliximab, golimumab, and the antibodies disclosed in U.S. Pat. No. 7,285,269, which is hereby incorporated by reference. Throughout this specification, “certolizumab” refers to an antibody having variable region sequences derived from certolizumab pegol. Preferred sequences based on such CDRs are shown in Table G. 
     
       
         
           
               
             
               
                 TABLE G 
               
             
            
               
                   
               
               
                 Preferred CDR sequences from anti-TNF-α antibodies 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 LCDR1 
                 LCDR2 
                 LCDR3 
                 HCDR1 
                 HCDR2 
                 HCDR3 
               
               
                 Antibody 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
                 (SEQ ID 
               
               
                 designation 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
                 NO) 
               
               
                   
               
               
                 adalimumab 
                 RASQGIRN 
                 AASTLQS 
                 QRYNRAPY 
                 DYAMH 
                 AITWNSGH 
                 VSYLSTAS 
               
               
                   
                 YLA (92) 
                 (242) 
                 T (96) 
                 (158) 
                 IDYADSVE 
                 SLDY (162) 
               
               
                   
                   
                   
                   
                   
                 G (160) 
                   
               
               
                   
               
               
                 certolizumab 
                 KASQNVGT 
                 SASFLYS 
                 QQYNIYPL 
                 DYGMN 
                 WINTYIGEP 
                 GYRSYAM 
               
               
                   
                 NVA (152) 
                 (154) 
                 T (156) 
                 (218) 
                 IYADSVKG 
                 DY (222) 
               
               
                   
                   
                   
                   
                   
                 (220) 
                   
               
               
                   
               
               
                 C234 
                 RASQDIRN 
                 AASSLQS 
                 LQHNSYPL 
                 SYDMH 
                 VISYDGSIK 
                 EVRSGSYY 
               
               
                   
                 DLG (140) 
                 (112) 
                 T (144) 
                 (206) 
                 YYADSVKG 
                 YYYSMDV 
               
               
                   
                   
                   
                   
                   
                 (208) 
                 (210) 
               
               
                   
               
               
                 3.2 
                 TGSSSNIG 
                 GNSNRPS 
                 QSYDSSLS 
                 SYWIG 
                 IIYLGDSDT 
                 SNWGLDY 
               
               
                   
                 AGYDVH 
                 (148) 
                 GSV (150) 
                 (212) 
                 RYSPSFQG 
                 (216) 
               
               
                   
                 (146) 
                   
                   
                   
                 (214) 
               
               
                   
               
            
           
         
       
     
     Each of the polypeptides in Table G is encoded by a nucleic acid having the sequence immediately preceding the sequence of the polypeptide in the Sequence Listing. The sequences in Table G may be used in any bispecific antigen binding protein format, preferably a hetero Ig or IgG-scFv format and preferably together with a TL1A antigen binding entity. 
     The TNF binding entities of the bispecific antigen binding proteins more preferably comprise the sequences of the variable region of selected anti-TNF antibodies. The sequences of such variable regions are set forth in Table H. As before, VL refers to the variable region of the light chain and VH refers to the variable region of the heavy chain. The invention further encompasses molecules having sequences at least about 90% identical to any of the sequences shown in Table H. 
     
       
         
           
               
             
               
                 TABLE H 
               
             
            
               
                   
               
               
                 Variable domain sequences of preferred anti-TNF-α antibodies 
               
            
           
           
               
               
            
               
                 Source 
                   
               
               
                 Antibody 
                   
               
               
                 designation 
                 Amino acid sequence (SEQ ID NO) 
               
               
                   
               
               
                 adalimumab 
                 DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGV 
               
               
                 VL 
                 PSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIK (2) 
               
               
                   
               
               
                 adalimumab 
                 EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNS 
               
               
                 VH 
                 GHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWG 
               
               
                   
                 QGTLVTVSS (4) 
               
               
                   
               
               
                 certolizuma 
                 DIQMTQSPSSLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPKALIYSASFLYSGV 
               
               
                 VL 
                 PYRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNIYPLTFGQGTKVEIK (42) 
               
               
                   
               
               
                 certolizumab 
                 EVQLVESGGGLVQPGGSLRLSCAASGYVFTDYGMNWVRQAPGKGLEWMGWINTYI 
               
               
                 VH 
                 GEPIYADSVKGRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCARGYRSYAMDYWGQG 
               
               
                   
                 TLVTVSS (44) 
               
               
                   
               
               
                 certolizumab 
                 DIQMTQSPSSLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPKALIYSASFLYSGV 
               
               
                 variant VL 
                 PYRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNIYPLTFGQGTKVEIK (42) 
               
               
                   
               
               
                 certolizumab 
                 EVQLVESGGGLVQPGGSLRLSCAASGYVFTDYGMNWVRQAPGKGLEWMGWINTYI 
               
               
                 variant VH 
                 GEPIYADSVKGRFTISLDTSKSTAYLQMNSLRAEDTAVYYCARGYRSYAMDYWGQGT 
               
               
                   
                 LVTVSS (318) 
               
               
                   
               
               
                 3.2 VL 
                 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQFPGTAPKLLIQGNSNRP 
               
               
                   
                 SGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVFGGGTKLTVL (38) 
               
               
                   
               
               
                 3.2 VH 
                 EVQLVQSGAEVKKPGESLKISCKTSEYSFTSYWIGWVRQMPGKGLEWMGIIYLGDSD 
               
               
                   
                 TRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARSNWGLDYWGQGTLVTV 
               
               
                   
                 SS (40) 
               
               
                   
               
               
                 C234 VL 
                 DIQMTQSPSSLSASVGDRVTITCRASQDIRNDLGWYQQKPGKAPKRLIYAASSLQSGV 
               
               
                   
                 PSRFSGSGSGPEFTLTISSLQPEDFATYYCLQHNSYPLTFGGGTKVEIK (34) 
               
               
                   
               
               
                 C234 VH 
                 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYDMHWVRQAPGKGLEWVAVISYDGS 
               
               
                   
                 IKYYADSVKGRFTISRDNSKNTLYLQVNSLRAEDTAVYYCAREVRSGSYYYYYSMDV 
               
               
                   
                 WGQGTTVTVSS (36) 
               
               
                   
               
            
           
         
       
     
     Each of the foregoing polypeptides is encoded by a nucleic acid having the nucleotide sequence immediately preceding the polypeptide sequence in the Sequence Listing. 
     Preferred anti-TL1A/anti-TNF hetero IgG bispecific antibodies are shown in Table I. The hetero Ig molecule designation is based on the foregoing designations for the parental antibodies used to construct the hetero Ig molecule. The molecule designated “certolizumab/3B3 variant,” for example, employs polypeptides having the sequences from antibodies designated certolizumab and 3B3 variant herein. As before, VL refers to the variable region of the light chain, VH to the variable region of the heavy chain. Such molecules may include a “C clamp,” cysteine molecules substituted into the sequences to provide an additional disulfide bond for stabilization. Such cysteine substitutions are preferably introduced at the VL-VH interface, at positions 44 in the VH and 100 in the VL in the Kabat numbering scheme. Additional substitutions may be made for stability or to introduce other functionality. Embodiments of the invention include molecules with at least 90% sequence identity to any of the sequences shown in Table I. 
     
       
         
           
               
             
               
                 TABLE I 
               
             
            
               
                   
               
               
                 Variable region sequences of anti-TL1A/ 
               
               
                 anti-TNF-α hetero IgG molecules 
               
            
           
           
               
               
               
            
               
                 Hetero Ig 
                 Anti-TNF-α binding entity 
                 Anti-TL1A binding entity 
               
            
           
           
               
               
               
               
               
            
               
                 molecule 
                 VL 
                 VH 
                 VL 
                 VH 
               
               
                 designation 
                 SEQ ID NO 
                 SEQ ID NO 
                 SEQ ID NO 
                 SEQ ID NO 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 Certolizumab/ 
                 42 
                 286 
                 288 
                 290 
               
               
                 3B3 variant 
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 42 
                 44 
                 292 
                 294 
               
               
                 2G11 variant 
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 42 
                 44 
                 296 
                 298 
               
               
                 23B3 VH4 
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 42 
                 44 
                 300 
                 302 
               
               
                 23B3 VH3 
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 42 
                 44 
                 304 
                 306 
               
               
                 3C6 variant 
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 42 
                 44 
                 308 
                 310 
               
               
                 3C6 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 288 
                 290 
               
               
                 3B3 variant 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 292 
                 294 
               
               
                 2G11 variant 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 296 
                 298 
               
               
                 23B3 VH4 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 300 
                 302 
               
               
                 23B3 VH3 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 304 
                 306 
               
               
                 3C6 variant 
                   
                   
                   
                   
               
               
                 3.2/ 
                 312 
                 314 
                 308 
                 310 
               
               
                 3C6 
                   
                   
                   
                   
               
               
                 Certolizumab 
                 42 
                 318 
                 288 
                 290 
               
               
                 variant/ 
                   
                   
                   
                   
               
               
                 3B3 variant 
                   
                   
                   
                   
               
               
                 Certolizumab 
                 42 
                 318 
                 292 
                 294 
               
               
                 variant/ 
                   
                   
                   
                   
               
               
                 2G11 variant 
                   
                   
                   
                   
               
               
                 Certolizumab 
                 42 
                 318 
                 296 
                 298 
               
               
                 variant/ 
                   
                   
                   
                   
               
               
                 23B3 VH4 
                   
                   
                   
                   
               
               
                 Certolizumab 
                 42 
                 318 
                 304 
                 306 
               
               
                 variant/ 
                   
                   
                   
                   
               
               
                 3C6 variant 
                   
                   
                   
                   
               
               
                 Certolizumab 
                 42 
                 318 
                 308 
                 310 
               
               
                 variant/ 
                   
                   
                   
                   
               
               
                 3C6 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 288 
                 290 
               
               
                 3B3 variant 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 292 
                 294 
               
               
                 2G11 variant 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 31 
                 298 
               
               
                 23B3 VH4 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 300 
                 302 
               
               
                 23B3 VH3 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 304 
                 306 
               
               
                 3C6 variant 
                   
                   
                   
                   
               
               
                 C234/ 
                 320 
                 322 
                 308 
                 310 
               
               
                 3C6 
                   
                   
                   
                   
               
               
                   
               
            
           
         
       
     
     Each of the foregoing polypeptides is encoded by a nucleic acid having the nucleotide sequence immediately preceding the amino acid sequence of the polypeptide in the Sequence Listing. 
     Table J lists the full light chain and heavy chain sequences of preferred hetero Ig bispecific antigen binding proteins in accordance with this invention. The listed sequences are preferred for all hetero Ig molecules, whether or not in the listed combination. The listed combinations are further preferred. “Var” in the molecule designation refers to variant. Such molecules may include charge variants to aid in assembly, as described for example in WO 2009/089004, published 16 Jul. 2009, hereby incorporated by reference. 
     
       
         
           
               
             
               
                 TABLE J 
               
             
            
               
                   
               
               
                 Full sequences of anti-TL1A/anti-TNF-α hetero IgG molecules 
               
            
           
           
               
               
               
               
            
               
                 Hetero Ig 
                 Anti-TNF-α binding entity 
                 Anti-TL1A binding entity 
                 Molecule 
               
            
           
           
               
               
               
               
               
               
            
               
                 molecule 
                 Light chain 
                 Heavy chain 
                 Light chain 
                 Heavy chain 
                 designation 
               
               
                 designation 
                 SEQ ID NO 
                 SEQ ID NO 
                 SEQ ID NO 
                 SEQ ID NO 
                 (iPS no.) 
               
               
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 134 
                 130 
                 340593 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 239 
                 138 
                 340595 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 253 
                 323 
                 340596 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 327 
                 325 
                 340597 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 331 
                 329 
                 340598 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 132 
                 136 
                 335 
                 329 
                 340599 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 134 
                 130 
                 340601 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 239 
                 138 
                 340602 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 323 
                 253 
                 340603 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 327 
                 325 
                 340604 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 331 
                 329 
                 340605 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 339 
                 335 
                 329 
                 340606 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 132 
                 316 
                 134 
                 130 
                 340607 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 132 
                 316 
                 239 
                 138 
                 340608 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 132 
                 316 
                 323 
                 253 
                 340609 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 132 
                 316 
                 335 
                 329 
                 340611 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 132 
                 316 
                 331 
                 329 
                 340610 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 316 
                 134 
                 130 
                 340612 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 316 
                 239 
                 138 
                 340613 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 316 
                 323 
                 253 
                 340614 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 316 
                 331 
                 329 
                 340615 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 316 
                 335 
                 329 
                 340616 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 463 
                 134 
                 130 
                 340617 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 463 
                 239 
                 138 
                 340618 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 463 
                 323 
                 253 
                 340619 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 463 
                 331 
                 329 
                 340620 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab var/ 
                 333 
                 463 
                 335 
                 329 
                 340621 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 134 
                 130 
                 349421 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 239 
                 138 
                 349425 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 323 
                 253 
                 349428 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 327 
                 325 
                 349431 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 331 
                 329 
                 349433 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 343 
                 335 
                 329 
                 349435 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 514 
                 512 
                 555 
                 349437 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 514 
                 541 
                 558 
                 349439 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 514 
                 539 
                 562 
                 349441 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 514 
                 543 
                 565 
                 349443 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 510 
                 514 
                 543 
                 568 
                 349445 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 510 
                 514 
                 544 
                 568 
                 349448 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 512 
                 555 
                 349451 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 541 
                 558 
                 349453 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 539 
                 562 
                 349455 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 543 
                 565 
                 349456 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 543 
                 568 
                 349457 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 542 
                 571 
                 544 
                 568 
                 349458 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 573 
                 512 
                 555 
                 349460 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 573 
                 541 
                 558 
                 349461 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 573 
                 539 
                 562 
                 349463 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 573 
                 543 
                 568 
                 349465 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 573 
                 544 
                 568 
                 349467 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 573 
                 512 
                 555 
                 349468 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 573 
                 541 
                 558 
                 349470 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 573 
                 539 
                 562 
                 349471 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 573 
                 543 
                 568 
                 349473 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 573 
                 544 
                 568 
                 349474 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 487 
                 512 
                 555 
                 349476 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 487 
                 541 
                 558 
                 349479 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 487 
                 539 
                 562 
                 349480 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 487 
                 543 
                 568 
                 349482 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 487 
                 544 
                 568 
                 349484 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 512 
                 555 
                 349485 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 541 
                 558 
                 349486 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 539 
                 562 
                 349487 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 543 
                 565 
                 349488 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 543 
                 568 
                 349489 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 544 
                 568 
                 349490 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 546 
                 579 
                 349491 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 548 
                 582 
                 349492 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 549 
                 585 
                 349493 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 550 
                 588 
                 349495 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 551 
                 591 
                 349496 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 545 
                 578 
                 552 
                 591 
                 349498 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 546 
                 579 
                 349499 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 548 
                 582 
                 349501 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 549 
                 585 
                 349502 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 550 
                 588 
                 349503 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 551 
                 591 
                 349504 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 547 
                 595 
                 552 
                 591 
                 349505 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 132 
                 599 
                 134 
                 598 
                 349506 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumb/ 
                 132 
                 599 
                 239 
                 601 
                 349507 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 132 
                 599 
                 323 
                 603 
                 349508 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 132 
                 599 
                 327 
                 605 
                 349509 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 132 
                 599 
                 331 
                 607 
                 349510 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 132 
                 599 
                 335 
                 607 
                 349511 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 134 
                 598 
                 349512 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 239 
                 601 
                 349513 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 323 
                 603 
                 349514 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 327 
                 605 
                 349515 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 331 
                 607 
                 349516 
               
               
                 3C6 variant 
                   
                   
                   
                   
                   
               
               
                 3.2/ 
                 337 
                 609 
                 335 
                 607 
                 349517 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 611 
                 134 
                 598 
                 349518 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 611 
                 239 
                 601 
                 349519 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 611 
                 323 
                 603 
                 349520 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 611 
                 331 
                 607 
                 349521 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 611 
                 335 
                 607 
                 349522 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 611 
                 134 
                 598 
                 349523 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 611 
                 239 
                 601 
                 349524 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 611 
                 323 
                 603 
                 349525 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 611 
                 331 
                 607 
                 349526 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 611 
                 335 
                 607 
                 349527 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 613 
                 134 
                 598 
                 349528 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 613 
                 239 
                 601 
                 349529 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 613 
                 323 
                 603 
                 349530 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 613 
                 335 
                 607 
                 349531 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 341 
                 615 
                 134 
                 598 
                 349532 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 615 
                 239 
                 601 
                 349533 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 615 
                 323 
                 603 
                 349534 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 615 
                 327 
                 605 
                 349535 
               
               
                 23B3 VH3 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 615 
                 331 
                 607 
                 349536 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 341 
                 615 
                 335 
                 607 
                 349537 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 333 
                 613 
                 331 
                 607 
                 349539 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 463 
                 239 
                 138 
                 361830 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 463 
                 331 
                 329 
                 361831 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 463 
                 335 
                 320 
                 361832 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 487 
                 512 
                 555 
                 361833 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 487 
                 541 
                 558 
                 361834 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 487 
                 539 
                 562 
                 361835 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 487 
                 543 
                 568 
                 361836 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 510 
                 487 
                 544 
                 568 
                 361837 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 613 
                 134 
                 598 
                 361838 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 613 
                 323 
                 603 
                 361839 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 613 
                 331 
                 607 
                 361840 
               
               
                 3C6 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 613 
                 335 
                 607 
                 361841 
               
               
                 3C6 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 132 
                 463 
                 134 
                 130 
                 361842 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 616 
                 512 
                 508 
                 381084 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 620 
                 512 
                 508 
                 381089 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 616 
                 541 
                 558 
                 381094 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 620 
                 541 
                 558 
                 381096 
               
               
                 2G11 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 616 
                 539 
                 562 
                 381098 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 540 
                 620 
                 539 
                 562 
                 381100 
               
               
                 23B3 VH4 
                   
                   
                   
                   
                   
               
               
                 C234 var/ 
                 538 
                 623 
                 512 
                 502 
                 381102 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 C234 var/ 
                 537 
                 626 
                 512 
                 508 
                 381109 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 546 
                 514 
                 194 
                 535 
                 381208 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 536 
                 616 
                 194 
                 535 
                 381216 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 536 
                 620 
                 194 
                 535 
                 381220 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 546 
                 487 
                 194 
                 535 
                 381222 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 471 
                 136 
                 473 
                 198 
                 381292 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 479 
                 477 
                 473 
                 198 
                 381300 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab/ 
                 176 
                 136 
                 520 
                 198 
                 381306 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 475 
                 477 
                 520 
                 198 
                 381312 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 471 
                 463 
                 473 
                 198 
                 381316 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 176 
                 463 
                 570 
                 198 
                 381318 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 471 
                 465 
                 473 
                 198 
                 381320 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 Certolizumab var/ 
                 176 
                 465 
                 520 
                 198 
                 381325 
               
               
                 3B3 var 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 514 
                 512 
                 508 
                 376541 
               
               
                 3B3 variant 
                   
                   
                   
                   
                   
               
               
                 C234/ 
                 518 
                 522 
                 512 
                 508 
                 376542 
               
               
                 3B3 variant 
                   
                   
                   
                   
                   
               
               
                 certolizumab/ 
                 510 
                 516 
                 512 
                 508 
                 376543 
               
               
                 3B3 variant 
                   
                   
                   
                   
                   
               
               
                   
               
            
           
         
       
     
     Each of the foregoing polypeptides is encoded by a nucleic acid having the nucleotide sequence immediately preceding the amino acid sequence of the polypeptide in the Sequence Listing, except as noted in Table J-1. 
     
       
         
           
               
             
               
                 TABLE J-1 
               
             
            
               
                   
               
               
                 Nucleic Acids Encoding Polypeptides of Table J 
               
            
           
           
               
               
               
            
               
                   
                 Polypeptide 
                 Encoding Nucleic Acid 
               
               
                   
                 SEQ ID NO: 
                 SEQ ID NO: 
               
               
                   
                   
               
               
                   
                 176 
                 526 
               
               
                   
                 194 
                 627 
               
               
                   
                 198 
                 529 
               
               
                   
                 239 
                 236 
               
               
                   
                 253 
                 250 
               
               
                   
                 320 
                 332 
               
               
                   
                 323 
                 257 
               
               
                   
                 333 
                 464 
               
               
                   
                 465 
                 528 
               
               
                   
                 471 
                 531 
               
               
                   
                 473 
                 530 
               
               
                   
                 475 
                 534 
               
               
                   
                 477 
                 532 
               
               
                   
                 479 
                 533 
               
               
                   
                 487 
                 515 
               
               
                   
                 508 
                 618 
               
               
                   
                 520 
                 527 
               
               
                   
                 535 
                 629 
               
               
                   
                 536 
                 630 
               
               
                   
                 537 
                 625 
               
               
                   
                 538 
                 622 
               
               
                   
                 539 
                 559 
               
               
                   
                 540 
                 560 
               
               
                   
                 541 
                 556 
               
               
                   
                 542 
                 570 
               
               
                   
                 543 
                 563 
               
               
                   
                 544 
                 569 
               
               
                   
                 545 
                 576 
               
               
                   
                 546 
                 574, 628 
               
               
                   
                 547 
                 593 
               
               
                   
                 548 
                 580 
               
               
                   
                 549 
                 583 
               
               
                   
                 550 
                 587 
               
               
                   
                 551 
                 590 
               
               
                   
                 552 
                 592 
               
               
                   
                 570 
                 527 
               
               
                   
                 571 
                 553 
               
               
                   
                 579 
                 575 
               
               
                   
                 588 
                 586 
               
               
                   
                 591 
                 589 
               
               
                   
                 598 
                 596 
               
               
                   
                 599 
                 597 
               
               
                   
                 616 
                 617 
               
               
                   
                 623 
                 621 
               
               
                   
                 626 
                 624 
               
               
                   
                   
               
            
           
         
       
     
     IgG-scFv Bispecific Antigen Binding Proteins 
     IgG-scFv antigen binding proteins have the structure shown in  FIG. 2 . Such molecules comprise the heavy and light chains of an antibody (the IgG portion of the molecule), which comprises a first antigen binding entity of the IgG-scFv molecule. Such molecules further comprise an scFv—a fusion protein having in one chain the heavy chain variable domain and light chain variable domain of a second antibody. This scFv portion of the molecule comprises a second antigen binding entity of the IgG-scFv molecule. An scFv portion is fused to each heavy chain of the IgG portion of the molecule. 
     An IgG-scFv antigen binding protein herein comprises an anti-TL1A binding entity, particularly the amino acid sequences of anti-TL1A binding entities as described hereinabove. Preferred IgG-scFv antigen binding proteins comprise the sequences of anti-TL1A CDRs as described in Tables A, B and C, with those of Table A most preferred. More preferred IgG-scFv antigen binding proteins comprise the anti-TL1A variable domain sequences of Table D. Also preferred are IgG-scFv antigen binding proteins having the full antibody sequences of Table E. 
     This specification further relates to IgG-scFv molecules comprising an anti-TNF-α antigen binding entity, preferably comprising CDR sequences as described in Table G. Further preferred are IgG-scFv molecules comprising the sequences of anti-TNF-α variable domains as described in Table H. Further preferred are IgG-scFv molecules comprising the full amino acid sequences of anti-TNF-α antibodies as described in Table K. 
     
       
         
           
               
             
               
                 TABLE K 
               
             
            
               
                   
               
               
                 Preferred anti-TNF-α Antibodies 
               
            
           
           
               
               
               
               
            
               
                 Antibody 
                 Light chain 
                 Heavy chain 
                   
               
               
                 designation 
                 SEQ ID NO 
                 SEQ ID NO 
                 iPS no. 
               
               
                   
               
               
                 Adalimumab 
                 46 
                 48 
                 104628 
               
               
                 C234 
                 78 
                 80 
                 330194 
               
               
                 3.2 
                 84 
                 86 
                 330237 
               
               
                 certolizumab 
                 88 
                 90 
                 333788 
               
               
                   
               
            
           
         
       
     
     Each of the amino acid sequences in Table K is encoded by the nucleic acid sequence immediately preceding it in the Sequence Listing. 
     In certain embodiments, the heavy chain of the IgG portion is fused to the scFv portion via a peptide linker. In certain embodiments, the peptide linker comprises a sequence selected from the group consisting of (Gly 3 Ser) 2 , (Gly 4 Ser) 2 , (Gly 3 Ser) 3 , (Gly 4 Ser) 3 , (Gly 3 Ser) 4 , (Gly 4 Ser) 4 , (Gly 3 Ser) 5 , (Gly 4 Ser) 5 , (Gly 3 Ser) 6 , and (Gly 4 Ser) 6 . 
     Further preferred are IgG-scFv antigen binding proteins having the heavy and light chain sequences as disclosed in Tables L and M. The heavy chain sequences in Tables L and M include the ScFv portion of the molecule fused to the heavy chain of the IgG portion of the molecule. In the molecule designation in the left column of these tables, the first antibody name identifies the IgG portion of the molecule, such that the IgG-scFv molecule comprises the full heavy and light chain sequences of the noted antibody. The second antibody name identifies the scFv portion of the molecule, such that the heavy chain sequence comprises the heavy chain variable domain and the light chain variable domain sequences as disclosed previously herein. Thus, “adalimumab IgG-9C8 scFv” includes the light chain of adalimumab and the heavy chain of adalimumab fused to heavy chain variable domain and light chain variable domain of antibody 9C8. “CC” in the molecule designations in Tables L and M refers to a cysteine clamp; molecules that include CC in their designation have been modified to have an additional cysteine disulfide bond. Such cysteine substitutions are preferably introduced at the VL-VH interface, at positions 44 in the VH and 100 in the VL in the Kabat numbering scheme. “Var” in Table L means variant. 
     
       
         
           
               
             
               
                 TABLE L 
               
             
            
               
                   
               
               
                 Preferred TL1A binding entity I2G-TNF binding entity scFv 
               
               
                 antigen binding proteins 
               
            
           
           
               
               
               
               
            
               
                   
                 heavy 
                 light 
                   
               
               
                   
                 chain 
                 chain 
                   
               
               
                   
                 SEQ ID 
                 SEQ ID 
                   
               
               
                 Molecule designation 
                 NO 
                 NO 
                 iPS 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 9C8 IgG-adalimumab scFv 
                 355 
                 447 
                 370012 
               
               
                 9C8 IgG-3.2 scFv 
                 357 
                 447 
                 370013 
               
               
                 9C8 IgG-C234 scFv 
                 359 
                 318 
                 370014 
               
               
                 9C8 IgG-adalimumab scFv + CC 
                 361 
                 447 
                 370015 
               
               
                 9C8 IgG-3.2 scFv + CC 
                 363 
                 447 
                 370016 
               
               
                 9C8 IgG-C234 scFv + CC 
                 365 
                 58 
                 370017 
               
               
                 23B3 VH4 IgG-adalimumab scFv 
                 367 
                 70 
                 370018 
               
               
                 23B3 VH4 IgG-3.2 scFv 
                 369 
                 70 
                 370019 
               
               
                 23B3 VH4 IgG-C234 scFv 
                 371 
                 70 
                 370020 
               
               
                 23B3 VH4 IgG-adalimumab scFV + CC 
                 373 
                 70 
                 370021 
               
               
                 23B3 VH4 IgG-3.2 scFv + CC 
                 375 
                 70 
                 370022 
               
               
                 23133 VH4 IgG-C234 scFv + CC 
                 377 
                 70 
                 370023 
               
               
                 3B3 var 2 IgG-adalimumab scFv 
                 391 
                 523 
                 371218 
               
               
                 3B3v2 IgG-3.2 scFv 
                 393 
                 523 
                 371219 
               
               
                 3B3v2 IgG-C234 scFv 
                 395 
                 523 
                 371220 
               
               
                 3B3v2 IgG-adalimumab scFv + CC 
                 397 
                 523 
                 371221 
               
               
                 3B3v2 IgG-3.2 scFv + CC 
                 399 
                 523 
                 371222 
               
               
                 3B3v2 IgG-C234 scFv + CC 
                 401 
                 523 
                 371223 
               
               
                 3B3 var IgG-3.2 var scFv 
                 403 
                 523 
                 381031 
               
               
                 3B3 var IgG-3.2 var scFv 
                 405 
                 523 
                 381035 
               
               
                 9C8 var IgG-3.2 var scFv 
                 407 
                 447 
                 381040 
               
               
                 9C8 var IgG-3.2 var scFv 
                 409 
                 447 
                 381044 
               
               
                 3B3 var IgG-3.2 var scFv + CC 
                 411 
                 523 
                 381048 
               
               
                 3B3 var IgG-3.2 var scFv + CC 
                 413 
                 523 
                 381053 
               
               
                 3B3 var IgG-C234 var scFv 
                 415 
                 523 
                 381058 
               
               
                 3B3 var IgG-C234 var scFv 
                 417 
                 523 
                 381062 
               
               
                 23B3 VH4 var IgG-adalimumab 
                 419 
                 70 
                 381066 
               
               
                 var scFv 
                   
                   
                   
               
               
                 23B3 VH4 var IgG-adalimumab 
                 421 
                 70 
                 381070 
               
               
                 var scFv 
                   
                   
                   
               
               
                 23B3 VH4 var IgG-adalimumab 
                 423 
                 70 
                 381074 
               
               
                 var scFv + CC 
                   
                   
                   
               
               
                 23B3 VH4 var IgG-adalimumab 
                 425 
                 70 
                 381079 
               
               
                 var scFv + CC 
               
               
                   
               
            
           
         
       
     
     Each of the polypeptides in Table L is encoded by a nucleic acid having the nucleotide sequence immediately preceding the polypeptide sequence in the Sequence Listing. 
     
       
         
           
               
             
               
                 TABLE M 
               
             
            
               
                   
               
               
                 Preferred TNF-α binding entity I2G-TL1A binding entity scFv 
               
               
                 antigen binding proteins 
               
            
           
           
               
               
               
               
            
               
                   
                 heavy 
                 light 
                   
               
               
                   
                 chain 
                 chain 
                   
               
               
                   
                 SEQ ID 
                 SEQ ID 
                   
               
               
                 Molecule designation 
                 NO 
                 NO 
                 iPS 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Adalimumab IgG-9C8 scFv 
                 82 
                 46 
                 369989 
               
               
                 Adalimumab IgG-23B3 VH4 scFv 
                 284 
                 46 
                 369990 
               
               
                 Adalimumab IgG-9C8 scFv + CC 
                 286 
                 46 
                 369992 
               
               
                 Adalimumab IgG-9C8 var scFv + CC 
                 441 
                 453 
                 381505 
               
               
                 Adalimumab IgG-23B3 VH4 scFv + CC 
                 312 
                 46 
                 369993 
               
               
                 3.2 IgG-9C8 scFv 
                 314 
                 84 
                 369995 
               
               
                 3.2 IgG-23B3 VH4 scFv 
                 320 
                 84 
                 369996 
               
               
                 3.2 IgG-9C8 scFv + CC 
                 322 
                 84 
                 369998 
               
               
                 3.2 IgG-23B3 VH4 scFv + CC 
                 345 
                 84 
                 369999 
               
               
                 C234 IgG-9C8 scFv 
                 347 
                 78 
                 370001  
               
               
                 C234 IgG-23B3 VH4 scFv 
                 349 
                 78 
                 370002 
               
               
                 C234 IgG-9C8 scFv + CC 
                 351 
                 78 
                 370004 
               
               
                 C234 IgG 23133 VH4 scFv + CC 
                 353 
                 78 
                 370005 
               
               
                 Adalimumab IgG-3B3v2 scFv 
                 379 
                 46 
                 371212 
               
               
                 Adalimumab IgG-3Bv2 scFv + CC 
                 381 
                 46 
                 371213 
               
               
                 3.2 IgG-3B3v2 scFv 
                 383 
                 84 
                 371214 
               
               
                 3.2 IgG-3B3v2 scFv + CC 
                 385 
                 84 
                 371215 
               
               
                 C234 IgG-3B3v2 scFv 
                 387 
                 78 
                 371216 
               
               
                 C234 IgG-3B3v2 scFv + CC 
                 389 
                 78 
                 371217 
               
               
                 3.2 var IgG-9C8 var scFv 
                 427 
                 449 
                 381327 
               
               
                 3.2 var IgG-9C8 var scFv 
                 429 
                 449 
                 381331 
               
               
                 C234 var IgG-9C8 var scFv 
                 431 
                 451 
                 381485 
               
               
                 C234 var IgG-9C8 var scFv + CC 
                 433 
                 451 
                 381489 
               
               
                 C234 var IgG-363 var scFv 
                 435 
                 451 
                 381493 
               
               
                 C234 var IgG-363 var scFv + CC 
                 437 
                 451 
                 381497 
               
               
                 Adalimumab var IgG-9C8 var scFv 
                 439 
                 453 
                 381501 
               
               
                 Adalimumab var IgG-9C8 var scFv + CC 
                 441 
                 453 
                 381505 
               
               
                 Adalimumab var IgG-3133 var scFv 
                 443 
                 453 
                 381509 
               
               
                 Adalimumab var IgG-363 var scFv + CC 
                 445 
                 453 
                 381513 
               
               
                   
               
            
           
         
       
     
     Each of the polypeptides in Table M is encoded by a nucleic acid having the nucleotide sequence immediately preceding the polypeptide sequence in the Sequence Listing. 
     IgG-Fab Bispecific Antigen Binding Proteins 
     In the IgG-Fab format, the bispecific, multivalent antigen binding protein comprises (i) a first polypeptide comprising a first heavy chain (VH2-CH1-CH2-CH3) from a first antibody, wherein the first heavy chain is fused at its carboxyl terminus (optionally through a peptide linker) to a polypeptide comprising VH2-CH1 domains of a second antibody to form a modified heavy chain, (ii) a second polypeptide comprising a light chain from a first antibody (VL1-CL) and (iii) a third polypeptide comprising VL2-CL domains of the second antibody. The CL and CH1 domains of the first antibody may be switched (“swapped”) in some embodiments between the first and second polypeptide. In such embodiments, the second polypeptide comprises VL1-CH1, while the first polypeptide comprises VH1-CL-CH2-CH3-VH2-CH1 with VH1-CL-CH2-CH3 fused at its C-terminus to VH2-CH1 optionally through a peptide linker. The third polypeptide comprises VL2-CL. Alternatively, the CL and CH1 domains of the second antibody may be switched in some embodiments between the first and third polypeptides. In such embodiments, the third polypeptide comprises VL2-CH1, while the first polypeptide comprises VH1-CH1-CH2-CH3-VH2-CL wherein VH1-CH1-CH2-CH1 is fused at its C-terminus to VH2-CL optionally through a peptide linker. The second polypeptide comprises VL1-CL. In yet another embodiment, the CL and CH1 domains of both antibodies are switched between the first, second and third polypeptides. In such embodiments, the first polypeptide comprises VH1-CL-CH2-CH3-VH2-CL, with VH1-CL-CH2-CH3 optionally fused at its C-terminus to VH2-CL, the second polypeptide comprises VL1-CH1, and the third polypeptide comprises VL2-CH1. Within the scope of this invention are such IgG-Fab molecules in which the first antibody comprises a TL1A binding entity and the second antibody comprises a TNF-α binding entity. Likewise, also within the scope of this invention are such IgG-Fab molecules in which the first antibody comprises a TNF-α binding entity and the second antibody comprises a TL1A binding entity. An expression having the same meaning as the last two sentences is that the first antibody comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In one aspect of the IgG-Fab format within this invention, the bispecific, tetravalent antigen binding protein comprises a) a first heavy chain of a first antibody (VH1), wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a moiety comprising a second heavy chain of a second antibody (VH2), wherein the second antibody specifically binds to a second antigen; b) two light chains of the first antibody of a); and c) two light chains of the second antibody of a). Within this invention, the first antibody may specifically bind TL1A and the second may specifically bind TNF-α or vice-versa. 
     In another aspect of the IgG-Fab format within this invention, the bispecific antigen binding protein comprises (i) a first binding domain that specifically binds to a first antigen comprising a first light chain immunoglobulin variable region (VL1) and a first heavy chain immunoglobulin variable region (VH1); (ii) a second binding domain that specifically binds to a second antigen comprising a second light chain immunoglobulin variable region (VL2) and a second heavy chain immunoglobulin variable region (VH2); and (iii) a human immunoglobulin Fc region, wherein one of the binding domains is positioned at the amino terminus of the Fc region and the other binding domain is positioned at the carboxyl terminus of the Fc region, wherein the carboxyl-terminal binding domain is a Fab and is fused through a peptide linker to the carboxyl terminus of the Fc region, and wherein the Fab is fused to the Fc region through the amino terminus of the VH region of the Fab. Within this invention, the first antigen may be TL1A and the second TNF-α or vice-versa. 
     In another aspect of the IgG-Fab format, the bispecific, tetravalent antigen binding protein comprises: 
     a) a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
         i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a charged (e.g., positively charged) amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a charged (preferably oppositely charged from that of i) above) amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering, wherein the charge is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain; and       

     b) a second polypeptide comprising a first light chain of the first antibody of a), wherein the first light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein the charge at position 38 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 183; and 
     c) a third polypeptide comprising a second light chain of the second antibody of a), wherein the second light chain comprises a second light chain variable region (VL2) and a second CL region; and wherein the VL2 or second CL domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein the charge at position 38 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 183. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In some embodiments of the bispecific antigen binding proteins of the invention in which the carboxyl-terminal binding domain is a Fab fragment, the binding domain positioned at the amino terminus of the Fc region (i.e., the amino-terminal binding domain) is also a Fab fragment. The amino-terminal Fab fragment can be fused to the amino terminus of the Fc region through a peptide linker or an immunoglobulin hinge region described herein. In some embodiments, the amino-terminal Fab fragment is joined to the amino terminus of the Fc region through a human IgG1 hinge region. In other embodiments, the amino-terminal Fab fragment is joined to the amino terminus of the Fc region through a human IgG2 hinge region. In one embodiment, the amino-terminal Fab fragment is fused to the Fc region through the carboxyl terminus of the CH1 region of the Fab. 
     In some embodiments, the bispecific antigen binding protein of the invention comprises a first antibody that specifically binds to a first target where one polypeptide chain (e.g. the heavy chain (VH2-CH1)) of a Fab fragment from a second antibody that specifically binds to a second target is fused to the carboxyl terminus of the heavy chain of the first antibody. The bispecific antigen binding protein in such embodiments also comprises a polypeptide chain containing the other half of the Fab fragment from the second antibody (e.g., the light chain (VL2-CL)). This format is referred to herein as the “IgG-Fab” format, and one embodiment of this type of molecule is shown schematically in  FIG. 25 . Thus, in certain embodiments, the present invention includes a bispecific, multivalent antigen binding protein comprising: (i) a light chain from a first antibody, (ii) a heavy chain from the first antibody, wherein the heavy chain is fused at its carboxyl terminus through a peptide linker to a first polypeptide comprising VH-CH1 domains of a second antibody to form a modified heavy chain, and (iii) a second polypeptide comprising VL-CL domains of the second antibody. When dimerized, the bispecific antigen binding protein is a homohexamer comprising two modified heavy chains, two light chains from the first antibody, and two polypeptide chains containing the other half of the Fab fragment from the second antibody (the Fd fragment). In one embodiment, the first polypeptide, which is fused to the carboxyl terminus of the heavy chain, comprises VH and CH1 domains from the second antibody, and the second polypeptide comprises VL and CL domains from the second antibody. 
     In certain embodiments, the antigen binding proteins of the invention comprise (i) a first binding domain that specifically binds a first target antigen, (ii) a second binding domain that specifically binds to a second target antigen, and (iii) a human immunoglobulin Fc region, wherein one of the binding domains is positioned at the amino terminus of the Fc region and the other binding domain is positioned at the carboxyl terminus of the Fc region. In some such embodiments, each of the first and second binding domains comprises immunoglobulin variable regions. For instance, in certain embodiments, the first binding domain comprises a first light chain variable region (VL1) and a first heavy chain variable region (VH1) from an anti-first target antigen antibody and the second binding domain comprises a second light chain variable region (VL2) and a second heavy chain variable region (VH2) from an anti-second target antigen antibody. 
     In certain embodiments of the bispecific antigen binding proteins of the invention, the binding domain positioned at the amino terminus of the Fc region (i.e. the amino-terminal binding domain) is a Fab fragment fused to the amino terminus of the Fc region through a peptide linker described herein or through an immunoglobulin hinge region. An “immunoglobulin hinge region” refers to the amino acid sequence connecting the CH1 domain and the CH2 domain of an immunoglobulin heavy chain. The hinge region of human IgG1 is generally defined as the amino acid sequence from about Glu216 or about Cys226, to about Pro230. Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain disulfide bonds in the same positions and are determinable to those of skill in the art. In some embodiments, the amino-terminal binding domain is joined to the amino terminus of the Fc region through a human IgG1 hinge region. In other embodiments, the amino-terminal binding domain is joined to the amino terminus of the Fc region through a human IgG2 hinge region. In one embodiment, the amino-terminal binding domain (e.g. Fab fragment) is fused to the Fc region through the carboxyl terminus of the CH1 region of the Fab. 
     In some embodiments of the antigen binding proteins of the invention, the binding domain positioned at the carboxyl terminus of the Fc region (i.e. the carboxyl-terminal binding domain) is a Fab fragment. In such embodiments, the Fab is fused or otherwise connected to the carboxyl terminus of the Fc region (e.g. the carboxyl terminus of the CH3 domain) through a peptide linker through the amino terminus of the VH region of the Fab fragment. Thus, in one embodiment, the Fab is fused to an Fc region through the amino terminus of the VH region of the Fab such that the resulting fusion protein comprises, from N-terminus to C-terminus, a CH2 domain, a CH3 domain, a peptide linker, a VH region, and a CH1 region. 
     In certain embodiments, the first heavy chain of an antigen binding protein of the present invention is fused to the VH2 via a peptide linker. In certain embodiments, the peptide linker comprises a sequence selected from the group consisting of (Gly 3 Ser) 2 , (Gly 4 Ser) 2 , (Gly 3 Ser) 3 , (Gly 4 Ser) 3 , (Gly 3 Ser) 4 , (Gly 4 Ser) 4 , (Gly 3 Ser) 5 , (Gly 4 Ser) 5 , (Gly 3 Ser) 6 , and (Gly 4 Ser) 6  These sequences can also be written as: 
     
       
         
           
               
               
            
               
                   
                 (SEQ ID NO: 673) 
               
               
                   
                 GGGSGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 695) 
               
               
                   
                 GGGGSGGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 703) 
               
               
                   
                 GGGSGGGSGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 729) 
               
               
                   
                 GGGGSGGGGSGGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 735) 
               
               
                   
                 GGGSGGGSGGGSGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 825) 
               
               
                   
                 GGGGSGGGGSGGGGSGGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 937) 
               
               
                   
                 GGGSGGGSGGGSGGGSGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 939) 
               
               
                   
                 GGGGSGGGGSGGGGSGGGGSGGGGS, 
               
               
                   
               
               
                   
                 (SEQ ID NO: 941) 
               
               
                   
                 GGGSGGGSGGGSGGGSGGGSGGGS, 
               
               
                   
                 and 
               
               
                   
               
               
                   
                 (SEQ ID NO: 945) 
               
               
                   
                 GGGGSGGGGSGGGGSGGGGSGGGGS. 
               
            
           
         
       
     
     The peptide linker joining the Fc region to the carboxyl-terminal Fab can be any of the peptide linkers described herein. In particular embodiments, the peptide linker joining the Fc region to the carboxyl-terminal Fab fragment is at least 5 amino acids in length. In other embodiments, the peptide linker joining the Fc region to the carboxyl-terminal Fab fragment is at least 8 amino acids in length. Particularly suitable peptide linkers for joining the Fc region to the carboxyl-terminal Fab fragment are glycine-serine linkers, such as (Gly x Ser) n  wherein x=3 or 4 and n=2, 3, 4, 5 or 6. In one embodiment, the peptide linker connecting the Fc region to the carboxyl-terminal Fab fragment is a L10 (G 4 S) 2  linker. In another embodiment, the peptide linker connecting the Fc region to the carboxyl-terminal Fab fragment is a L9 or G 3 SG 4 S linker (GGGSGGGGS, SEQ ID NO: 946). 
     Chain Mutations 
     In particular embodiments, the bispecific antigen binding proteins described herein comprise a Fc region from a human IgG1 antibody modified to remove effector functions. Such modified IgG1 Fc regions comprise substitutions R292C and V302C. In such embodiments, the Fc region may also comprise substitution N297G (known as a SEFL2 scaffold), 
     Preferred embodiments further include mutations in the heavy and light chains to aid in correct assembly of a hetero IgG bispecific antigen binding protein. An approach for promoting heterodimer formation to the exclusion of homodimer formation entails utilizing an electrostatic steering mechanism (see Gunasekaran et al. (2010),  J. Biol. Chem.,  Vol. 285: 19637-19646, which is hereby incorporated by reference in its entirety). This approach involves introducing or exploiting charged residues in the CH3 domain in each heavy chain so that the two different heavy chains associate through opposite charges that cause electrostatic attraction. Homodimerization of the identical heavy chains are disfavored because the identical heavy chains have the same charge and therefore are repelled. This same electrostatic steering technique can be used to prevent mispairing of light chains with the non-cognate heavy chains by introducing residues having opposite charges in the correct light chain—heavy chain pair at the binding interface. The electrostatic steering technique and suitable charge pair mutations for promoting heterodimers and correct light chain-heavy chain pairing is described in WO2009/089004 and WO2014/081955, both of which are hereby incorporated by reference in their entireties. 
     In embodiments in which the bispecific antigen binding proteins of the invention are heterodimeric antibodies comprising a first light chain (LC1) and first heavy chain (HC1) from a first antibody that specifically binds to a first target antigen and a second light chain (LC2) and second heavy chain (HC2) from a second antibody that specifically binds to a second target, HC1 or HC2 may comprise one or more amino acid substitutions to replace a positively-charged amino acid with a negatively-charged amino acid. For instance, in one embodiment, the CH3 domain of HC1 or the CH3 domain of HC2 comprises an amino acid sequence differing from a wild-type IgG amino acid sequence such that one or more positively-charged amino acids (e.g., lysine, histidine and arginine) in the wild-type human IgG amino acid sequence are replaced with one or more negatively-charged amino acids (e.g., aspartic acid and glutamic acid) at the corresponding position(s) in the CH3 domain. In these and other embodiments, amino acids (e.g. lysine) at one or more positions selected from 370, 392 and 409 (EU numbering system) are replaced with a negatively-charged amino acid (e.g., aspartic acid and glutamic acid). An amino acid substitution in an amino acid sequence is typically designated herein with a one-letter abbreviation for the amino acid residue in a particular position, followed by the numerical amino acid position relative to an original sequence of interest, which is then followed by the one-letter symbol for the amino acid residue substituted in. For example, “T3OD” symbolizes a substitution of a threonine residue by an aspartate residue at amino acid position 30, relative to the original sequence of interest. Another example, “S218G” symbolizes a substitution of a serine residue by a glycine residue at amino acid position 218, relative to the original amino acid sequence of interest. 
     In certain embodiments, HC1 or HC2 of the heterodimeric antibodies may comprise one or more amino acid substitutions to replace a negatively-charged amino acid with a positively-charged amino acid. For instance, in one embodiment, the CH3 domain of HC1 or the CH3 domain of HC2 comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more negatively-charged amino acids in the wild-type human IgG amino acid sequence are replaced with one or more positively-charged amino acids at the corresponding position(s) in the CH3 domain. In these and other embodiments, amino acids (e.g., aspartic acid or glutamic acid) at one or more positions selected from 356, 357, and 399 (EU numbering system) of the CH3 domain are replaced with a positively-charged amino acid (e.g., lysine, histidine and arginine). 
     In particular embodiments, the heterodimeric antibody comprises a first heavy chain comprising negatively-charged amino acids at positions 392 and 409 (e.g., K392D and K409D substitutions), and a second heavy chain comprising positively-charged amino acids at positions 356 and 399 (e.g., E356K and D399K substitutions). In other particular embodiments, the heterodimeric antibody comprises a first heavy chain comprising negatively-charged amino acids at positions 392, 409, and 370 (e.g., K392D, K409D, and K370D substitutions), and a second heavy chain comprising positively-charged amino acids at positions 356, 399, and 357 (e.g., E356K, D399K, and E357K substitutions). In related embodiments, the first heavy chain is from an anti-first target antigen antibody and the second heavy chain is from an anti-second target antigen antibody. 
     To facilitate the association of a particular heavy chain with its cognate light chain, both the heavy and light chains may contain complementary amino acid substitutions. As used herein, “complementary amino acid substitutions” refer to a substitution to a positively-charged amino acid in one chain paired with a negatively-charged amino acid substitution in the other chain. For example, in some embodiments, the heavy chain comprises at least one amino acid substitution to introduce a charged amino acid and the corresponding light chain comprises at least one amino acid substitution to introduce a charged amino acid, wherein the charged amino acid introduced into the heavy chain has the opposite charge of the amino acid introduced into the light chain. In certain embodiments, one or more positively-charged residues (e.g., lysine, histidine or arginine) can be introduced into a first light chain (LC1) and one or more negatively-charged residues (e.g., aspartic acid or glutamic acid) can be introduced into the companion heavy chain (HC1) at the binding interface of LC1/HC1, whereas one or more negatively-charged residues (e.g., aspartic acid or glutamic acid) can be introduced into a second light chain (LC2) and one or more positively-charged residues (e.g., lysine, histidine or arginine) can be introduced into the companion heavy chain (HC2) at the binding interface of LC2/HC2. The electrostatic interactions will direct the LC1 to pair with HC1 and LC2 to pair with HC2, as the opposite charged residues (polarity) at the interface attract. The heavy/light chain pairs having the same charged residues (polarity) at an interface (e.g. LC1/HC2 and LC2/HC1) will repel, resulting in suppression of the unwanted HC/LC pairings. 
     In these and other embodiments, the CH1 domain of the heavy chain or the CL domain of the light chain comprises an amino acid sequence differing from a wild-type IgG amino acid sequence such that one or more positively-charged amino acids in a wild-type IgG amino acid sequence is replaced with one or more negatively-charged amino acids. Alternatively, the CH1 domain of the heavy chain or the CL domain of the light chain comprises an amino acid sequence differing from a wild-type IgG amino acid sequence such that one or more negatively-charged amino acids in a wild-type IgG amino acid sequence is replaced with one or more positively-charged amino acids. In some embodiments, one or more amino acids in the CH1 domain of the first and/or second heavy chain in the heterodimeric antibody at an EU position selected from F126, P127, L128, A141, L145, K147, D148, H168, F170, P171, V173, Q175, S176, S183, V185 and K213 is replaced with a charged amino acid. In certain embodiments, a heavy chain residue for substitution with a negatively- or positively-charged amino acid is 5183 (EU numbering system). In some embodiments, 5183 is substituted with a positively-charged amino acid. In alternative embodiments, S183 is substituted with a negatively-charged amino acid. For instance, in one embodiment, S183 is substituted with a negatively-charged amino acid (e.g. S183E) in the first heavy chain, and 5183 is substituted with a positively-charged amino acid (e.g. S183K) in the second heavy chain. 
     In embodiments in which the light chain is a kappa light chain, one or more amino acids in the CL domain of the first and/or second light chain in the heterodimeric antibody at a position (EU numbering in a kappa light chain) selected from F116, F118, S121, D122, E123, Q124, S131, V133, L135, N137, N138, Q160, S162, T164, S174 and S176 is replaced with a charged amino acid. In embodiments in which the light chain is a lambda light chain, one or more amino acids in the CL domain of the first and/or second light chain in the heterodimeric antibody at a position (EU numbering in a lambda chain) selected from T116, F118, S121, E123, E124, K129, T131, V133, L135, S137, E160, T162, S165, Q167, A174, S176 and Y178 is replaced with a charged amino acid. In some embodiments, a residue for substitution with a negatively- or positively-charged amino acid is S176 (EU numbering system) of the CL domain of either a kappa or lambda light chain. In certain embodiments, S176 of the CL domain is replaced with a positively-charged amino acid. In alternative embodiments, S176 of the CL domain is replaced with a negatively-charged amino acid. In one embodiment, S176 is substituted with a positively-charged amino acid (e.g. S176K) in the first light chain, and S176 is substituted with a negatively-charged amino acid (e.g. S176E) in the second light chain. 
     In addition to or as an alternative to the complementary amino acid substitutions in the CH1 and CL domains, the variable regions of the light and heavy chains in the heterodimeric antibody may contain one or more complementary amino acid substitutions to introduce charged amino acids. For instance, in some embodiments, the VH region of the heavy chain or the VL region of the light chain of a heterodimeric antibody comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more positively-charged amino acids in wild-type IgG amino acid sequence is replaced with one or more negatively-charged amino acids. Alternatively, the VH region of the heavy chain or the VL region of the light chain comprises an amino acid sequence differing from a wild-type IgG amino acid sequence such that one or more negatively-charged amino acids in a wild-type IgG amino acid sequence is replaced with one or more positively-charged amino acids. 
     V region interface residues (i.e., amino acid residues that mediate assembly of the VH and VL regions) within the VH region include EU positions 1, 3, 35, 37, 39, 43, 44, 45, 46, 47, 50, 59, 89, 91, and 93. One or more of these interface residues in the VH region can be substituted with a charged (positively- or negatively-charged) amino acid. In certain embodiments, the amino acid at EU position 39 in the VH region of the first and/or second heavy chain is substituted for a positively-charged amino acid, e.g., lysine. In alternative embodiments, the amino acid at EU position 39 in the VH region of the first and/or second heavy chain is substituted for a negatively-charged amino acid, e.g., glutamic acid. In some embodiments, the amino acid at EU position 39 in the VH region of the first heavy chain is substituted for a negatively-charged amino acid (e.g., G39E), and the amino acid at EU position 39 in the VH region of the second heavy chain is substituted for a positively-charged amino acid (e.g., G39K). In some embodiments, the amino acid at EU position 44 in the VH region of the first and/or second heavy chain is substituted for a positively-charged amino acid; for example, lysine. In alternative embodiments, the amino acid at EU position 44 in the VH region of the first and/or second heavy chain is substituted for a negatively-charged amino acid; for example, glutamic acid. In certain embodiments, the amino acid at EU position 44 in the VH region of the first heavy chain is substituted for a negatively-charged amino acid (e.g., G44E), and the amino acid at EU position 44 in the VH region of the second heavy chain is substituted for a positively-charged amino acid (e.g., G44K). 
     V region interface residues (i.e., amino acid residues that mediate assembly of the VH and VL regions) within the VL region include EU positions 32, 34, 35, 36, 38, 41, 42, 43, 44, 45, 46, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 85, 87, 89, 90, 91, and 100. One or more interface residues in the VL region can be substituted with a charged amino acid, preferably an amino acid that has an opposite charge to those introduced into the VH region of the cognate heavy chain. In some embodiments, the amino acid at EU position 100 in the VL region of the first and/or second light chain is substituted for a positively-charged amino acid; for example, lysine. In alternative embodiments, the amino acid at EU position 100 in the VL region of the first and/or second light chain is substituted for a negative-charged amino acid; for example,s glutamic acid. In certain embodiments, the amino acid at EU position 100 in the VL region of the first light chain is substituted for a positively-charged amino acid (e.g. G100K), and the amino acid at EU position 100 in the VL region of the second light chain is substituted for a negatively-charged amino acid (e.g. G100E). 
     In certain embodiments, a heterodimeric antibody of the invention comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the first heavy chain comprises amino acid substitutions at positions 44 (EU), 183 (EU), 392 (EU) and 409 (EU), wherein the second heavy chain comprises amino acid substitutions at positions 44 (EU), 183 (EU), 356 (EU) and 399 (EU), wherein the first and second light chains comprise an amino acid substitution at positions 100 (EU) and 176 (EU), and wherein the amino acid substitutions introduce a charged amino acid at the positions. In related embodiments, the glycine at position 44 (EU) of the first heavy chain is replaced with glutamic acid, the glycine at position 44 (EU) of the second heavy chain is replaced with lysine, the glycine at position 100 (EU) of the first light chain is replaced with lysine, the glycine at position 100 (EU) of the second light chain is replaced with glutamic acid, the serine at position 176 (EU) of the first light chain is replaced with lysine, the serine at position 176 (EU) of the second light chain is replaced with glutamic acid, the serine at position 183 (EU) of the first heavy chain is replaced with glutamic acid, the lysine at position 392 (EU) of the first heavy chain is replaced with aspartic acid, the lysine at position 409 (EU) of the first heavy chain is replaced with aspartic acid, the serine at position 183 (EU) of the second heavy chain is replaced with lysine, the glutamic acid at position 356 (EU) of the second heavy chain is replaced with lysine, and/or the aspartic acid at position 399 (EU) of the second heavy chain is replaced with lysine. 
     In other embodiments, a heterodimeric antibody of the invention comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the first heavy chain comprises amino acid substitutions at positions 183 (EU), 392 (EU) and 409 (EU), wherein the second heavy chain comprises amino acid substitutions at positions 183 (EU), 356 (EU) and 399 (EU), wherein the first and second light chains comprise an amino acid substitution at position 176 (EU), and wherein the amino acid substitutions introduce a charged amino acid at the positions. In related embodiments, the serine at position 176 (EU) of the first light chain is replaced with lysine, the serine at position 176 (EU) of the second light chain is replaced with glutamic acid, the serine at position 183 (EU) of the first heavy chain is replaced with glutamic acid, the lysine at position 392 (EU) of the first heavy chain is replaced with aspartic acid, the lysine at position 409 (EU) of the first heavy chain is replaced with aspartic acid, the serine at position 183 (EU) of the second heavy chain is replaced with lysine, the glutamic acid at position 356 (EU) of the second heavy chain is replaced with lysine, and/or the aspartic acid at position 399 (EU) of the second heavy chain is replaced with lysine. 
     In still other embodiments, a heterodimeric antibody of the invention comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the first heavy chain comprises amino acid substitutions at positions 183 (EU), 392 (EU), 409 (EU), and 370 (EU), wherein the second heavy chain comprises amino acid substitutions at positions 183 (EU), 356 (EU), 399 (EU), and 357 (EU), wherein the first and second light chains comprise an amino acid substitution at position 176 (EU), and wherein the amino acid substitutions introduce a charged amino acid at the positions. In related embodiments, the serine at position 176 (EU) of the first light chain is replaced with lysine, the serine at position 176 (EU) of the second light chain is replaced with glutamic acid, the serine at position 183 (EU) of the first heavy chain is replaced with glutamic acid, the lysine at position 392 (EU) of the first heavy chain is replaced with aspartic acid, the lysine at position 409 (EU) of the first heavy chain is replaced with aspartic acid, the lysine at position 370 (EU) of the first heavy chain is replaced with aspartic acid, the serine at position 183 (EU) of the second heavy chain is replaced with lysine, the glutamic acid at position 356 (EU) of the second heavy chain is replaced with lysine, the aspartic acid at position 399 (EU) of the second heavy chain is replaced with lysine, and/or the glutamic acid at position 357 (EU) of the second heavy chain is replaced with lysine. 
     Any of the constant domains can be modified to contain one or more of the charge pair mutations described above to facilitate correct assembly of a heterodimeric antibody. 
     The inventive heterodimeric antibodies also encompass antibodies comprising the heavy chain(s) and/or light chain(s), where one, two, three, four or five amino acid residues are lacking from the N-terminus or C-terminus, or both, in relation to any one of the heavy and light chains, e.g., due to post-translational modifications resulting from the type of host cell in which the antibodies are expressed. For instance, Chinese Hamster Ovary (CHO) cells frequently cleave off a C-terminal lysine from antibody heavy chains. 
     Charge pair mutations or complementary amino acid substitutions as described herein can be introduced into the Fab regions of the first antibody (Fab 1) or second antibody (Fab 2) to promote correct heavy chain-light chain pairing. For instance, in some embodiments, the amino acid at EU position 38 of the VL domain in Fab 1 is replaced with a negatively-charged amino acid (e.g. glutamic acid) and the amino acid at EU position 39 of the VH domain in Fab 1 is replaced with a positively-charged amino acid (e.g. lysine). In other embodiments, the amino acid at EU position 38 of the VL domain in Fab 1 is replaced with a positively-charged amino acid (e.g. lysine) and the amino acid at EU position 39 of the VH domain in Fab 1 is replaced with a negatively-charged amino acid (e.g. glutamic acid). In certain embodiments, the amino acid at EU position 38 of the VL domain in Fab 2 is replaced with a negatively-charged amino acid (e.g. glutamic acid) and the amino acid at EU position 39 of the VH domain in Fab 2 is replaced with a positively-charged amino acid (e.g. lysine). In other embodiments, the amino acid at EU position 38 of the VL domain in Fab 2 is replaced with a positively-charged amino acid (e.g. lysine) and the amino acid at EU position 39 of the VH domain in Fab 2 is replaced with a negatively-charged amino acid (e.g. glutamic acid). 
     In embodiments in which the VH-CH1 region (i.e. Fd fragment) from the second antibody is fused to the heavy chain of the first antibody, the heavy chain from the first antibody comprises a S183E mutation (EU numbering), the light chain from the first antibody comprises a S176K mutation (EU numbering), the light chain from the second antibody comprises a S176E mutation (EU numbering), and the Fd region from the second antibody (which is fused to the C-terminus of the heavy chain from the first antibody) comprises a S183K mutation (EU numbering). In other embodiments, the heavy chain from the first antibody comprises a G44E mutation (EU) and S183E mutation (EU numbering), the light chain from the first antibody comprises a G100K mutation (EU) and S176K mutation (EU numbering), the light chain from the second antibody comprises a G100E mutation (EU) and S176E mutation (EU numbering), and the Fd region from the second antibody (which is fused to the C-terminus of the heavy chain from the first antibody) comprises a G44K mutation (EU) and S183K mutation (EU numbering). The charges in the foregoing examples may be reversed so long as the charge on the corresponding light or heavy chain is also reversed so that the correct heavy/light chain pairs have opposite charges. 
     In one embodiment, the present invention is directed to a bispecific, tetravalent antigen binding protein, comprising: 
     a) a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
         i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a charged (e.g., positively charged) amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce an oppositely charged (e.g., negatively charged) amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering; and       

     b) a second polypeptide comprising a first light chain of the first antibody of a), wherein the first light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering; and 
     c) a third polypeptide comprising a second light chain of the second antibody of a), wherein the second light chain comprises a second light chain variable region (VL2) and a second CL region;. and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     “Corresponds to” as it pertains to the VH2 and second CH1 domain means that the amino acid residues of the VH2 and second CH1 domain are counted from the C-terminus of the first heavy chain if there is no linker. If there is a peptide linker, the amino acid residues of the VH2 and second CH1 domain are counted from the C-terminus of the peptide linker. In neither case are the amino acid residues counted from the N-terminus of the first heavy chain. Rather, for the VH2 and second CH1 domain, counting begins at the first amino acid residue of the VH2 domain. The counting of amino acid residues is performed using the EU or AHo convention. 
     In certain embodiments: a) the VH1 or first CH1 domain comprises a mutation selected from the group consisting of Q39K, G44K, and S183K using EU numbering; b) the VH2 or second CH1 domain comprises a mutation selected from the group consisting of Q39E, G44E, and S183E using EU numbering; c) the VL1 or first CL domain comprises a mutation selected from the group consisting of Q38E, G100E, and S176E using EU numbering; and d) the VL2 or second CL domain comprises a mutation selected from the group consisting of Q38K, G100K, and S176K using EU numbering. 
     In certain embodiments: a) the VH1 comprises a Q39K mutation and the first CH1 domain comprises a S183K mutation using EU numbering; b) the VH2 comprises a Q39E mutation and the second CH1 domain comprises a S183E mutation using EU numbering; c) the VL1 comprises a Q38E mutation and the first CL domain comprises a S176E mutation using EU numbering; and d) the VL2 comprises a Q38K mutation and the second CL domain comprises a S176K mutation using EU numbering. 
     In certain embodiments: a) the first CH1 domain comprises G44K and S183K mutations using EU numbering; b) the second CH1 domain comprises G44E and S183E mutations using EU numbering; c) the first CL domain comprises G100E and S176E mutations using EU numbering; and d) the second CL domain comprises G100K and S176K mutations using EU numbering. 
     In one embodiment the present invention is directed to a bispecific, tetravalent antigen binding protein, comprising: 
     a) a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
         i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering; and       

     b) a second polypeptide comprising a first light chain of the first antibody of a), wherein the first light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering; and 
     c) a third polypeptide comprising a second light chain of the second antibody of a), wherein the second light chain comprises a second light chain variable region (VL2) and a second CL region;. and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In certain embodiments: a) the VH1 or first CH1 domain comprises a mutation selected from the group consisting of Q39E, G44E, and S183E using EU numbering; b) the VH2 or second CH1 domain comprises a mutation selected from the group consisting of Q39K, G44K, and S183K using EU numbering; c) the VL1 or first CL domain comprises a mutation selected from the group consisting of Q38K, G100K, and S176K using EU numbering; and d) the VL2 or second CL domain comprises a mutation selected from the group consisting of Q38E, G100E, and S176E using EU numbering. 
     In certain embodiments: a) the first CH1 domain comprises a S183E mutation using EU numbering; b) the second CH1 domain comprises a S183K mutation using EU numbering; c) the first CL domain comprises a S176K mutation using EU numbering; and d) the second CL domain comprises a S176E mutation using EU numbering. 
     In certain embodiments: a) the VH1 comprises a Q39E mutation and the first CH1 domain comprises a S183E mutation using EU numbering; b) the VH2 comprises a Q39K mutation and the second CH1 domain comprises a S183K mutation using EU numbering; c) the VL1 comprises a Q38K mutation and the first CL domain comprises a S176K mutation using EU numbering; and d) the VL2 comprises a Q38E mutation and the second CL domain comprises a S176E mutation using EU numbering. 
     In certain embodiments: a) the first CH1 domain comprises G44E and S183E mutations using EU numbering; b) the second CH1 domain comprises G44K and S183K mutations using EU numbering; c) the first CL domain comprises G100K and S176K mutations using EU numbering; and d) the second CL domain comprises G100E and S176E mutations using EU numbering. 
     In one embodiment the present invention is directed to a bispecific, tetravalent antigen binding protein, comprising: 
     a) a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
         i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering, wherein the charge is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain; and       

     b) a second polypeptide comprising a first light chain of the first antibody of a), wherein the first light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein 
     the charge at position 38 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 183; and 
     c) a third polypeptide comprising a second light chain of the second antibody of a), wherein the second light chain comprises a second light chain variable region (VL2) and a second CL region; and wherein the VL2 or second CL domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein 
     the charge at position 38 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 183. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In certain embodiments: a) the VH1 comprises a Q39E mutation and the first CH1 domain comprises a S183K mutation using EU numbering; b) the VH2 comprises a Q39K mutation and the second CH1 domain comprises a S183E mutation using EU numbering; c) the VL1 comprises a Q38K mutation and the first CL domain comprises a S176E mutation using EU numbering; and d) the VL2 comprises a Q38E mutation and the second CL domain comprises a S176K mutation using EU numbering. 
     In certain embodiments: a) the first CH1 domain comprises G44E and S183K mutations using EU numbering; b) the second CH1 domain comprises G44K and S183E mutations using EU numbering; c) the first CL domain comprises G100K and S176E mutations using EU numbering; and d) the second CL domain comprises G100E and S176K mutations using EU numbering. 
     In certain embodiments: a) the VH1 comprises a Q39K mutation and the first CH1 domain comprises a S183E mutation using EU numbering; b) the VH2 comprises a Q39E mutation and the second CH1 domain comprises a S183K mutation using EU numbering; c) the VL1 comprises a Q38E mutation and the first CL domain comprises a S176K mutation using EU numbering; and d) the VL2 comprises a Q38K mutation and the second CL domain comprises a S176E mutation using EU numbering. 
     In certain embodiments: a) the first CH1 domain comprises G44K and S183E mutations using EU numbering; b) the second CH1 domain comprises G44E and S183K mutations using EU numbering; c) the first CL domain comprises G100E and S176K mutations using EU numbering; and d) the second CL domain comprises G100K and S176E mutations using EU numbering. 
     In one embodiment the present invention is directed to a method for preparing a bispecific, tetravalent antigen binding protein, comprising: 
     1) co-expressing in a host cell;
         a) a first polynucleotide wherein the first polynucleotide encodes a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
           i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering; and   
           b) a second polynucleotide wherein the second polynucleotide encodes a second polypeptide comprising a light chain of the first antibody of a), wherein the light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering;   c) a third polynucleotide wherein the third polycucleotide encodes a third polypeptide comprising a light chain of the second antibody of a), wherein the light chain comprises a second light chain variable region (VL2) and a second CL region;. and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering;       

     2) cultivating the host cell under conditions such that the polypeptides are produced; and 
     3) recovering from the host cell the antigen binding protein. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In one embodiment the present invention is directed to a method for preparing a bispecific, tetravalent antigen binding protein, comprising: 
     1) co-expressing in a host cell:
         a) a first polynucleotide wherein the first polynucleotide encodes a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
           i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering; and   
           b) a second polynucleotide wherein the second polynucleotide encodes a second polypeptide comprising a first light chain of the first antibody of a), wherein the first light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering; and   c) a third polynucleotide wherein the third polynucleotide encodes a third polypeptide comprising a second light chain of the second antibody of a), wherein the second light chain comprises a second light chain variable region (VL2) and a second CL region;. and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a negatively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering;       

     2) cultivating the host cell under conditions such that the polypeptides are produced; and 
     3) recovering from the host cell the antigen binding protein. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     In one embodiment the present invention is directed to a method for preparing a bispecific, tetravalent antigen binding protein, comprising: 
     1) co-expressing in a host cell:
         a) a first polynucleotide wherein the first polynucleotide encodes a first polypeptide comprising a first heavy chain of a first antibody comprising a first heavy chain variable region (VH1) and a first CH1 domain, wherein the first antibody specifically binds to a first antigen, and wherein the first heavy chain is fused through its C-terminus to the N-terminus of a polypeptide comprising a second heavy chain variable region of a second antibody (VH2), wherein the VH2 is fused through its C-terminus to the N-terminus of a second CH1 domain, and wherein the second antibody specifically binds to a second antigen; wherein
           i) the VH1 or first CH1 domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 39, 44, and 183 using EU numbering; and   ii) the VH2 or second CH1 domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of a residue that corresponds to positions 39, 44, and 183 using EU numbering, wherein the charge is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain; and   
           b) a second polynucleotide wherein the second polynucleotide encodes a second polypeptide comprising a light chain of the first antibody of a), wherein the light chain comprises a first light chain variable region (VL1) and a first CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein
           the charge at position 38 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH1 or first CH1 of the first heavy chain at position 183; and   
           c) a third polynucleotide wherein the third polynucleotide encodes a third polypeptide comprising a light chain of the second antibody of a), wherein the light chain comprises a second light chain variable region (VL2) and a second CL region; and wherein the VL1 or first CL domain comprises at least one amino acid substitution to introduce a positively charged amino acid at a residue selected from the group consisting of positions 38, 100, and 176 using EU numbering, wherein
           the charge at position 38 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 39; the charge at position 100 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 44; the charge at position 176 is the opposite of the substituted residue of the VH2 or second CH1 of the second heavy chain at position 183;   
               

     2) cultivating the host cell under conditions such that the polypeptides are produced; and 
     3) recovering from the host cell the antigen binding protein. 
     Within this invention, the first antigen in subparagraph a) above may be one of TL1A and TNF-α and the second antigen may be the other. Another way of stating this same binding specificity is to say that the first antibody in subparagraph a) comprises one of a TL1A binding entity or a TNF-α binding entity and the second antibody comprises the other. 
     Additionally or alternatively, correct heavy-light chain pairing may be facilitated by swapping the CH1 and CL domains in the carboxyl-terminal Fab binding domain. By way of example, the first polypeptide, which is fused to the carboxyl terminus of the heavy chain, may comprise a VL domain and CH1 domain from the second antibody, and the second polypeptide may comprise a VH domain and CL domain from the second antibody. In another embodiment, the first polypeptide, which is fused to the carboxyl terminus of the heavy chain, may comprise a VH domain and a CL domain from the second antibody, and the second polypeptide may comprise a VL domain and CH1 domain from the second antibody. 
     The heavy chain constant regions or the Fc regions of the bispecific antigen binding proteins described herein may comprise one or more amino acid substitutions that affect the glycosylation and/or effector function of the antigen binding protein. One of the functions of the Fc region of an immunoglobulin is to communicate to the immune system when the immunoglobulin binds its target. This is commonly referred to as “effector function.” Communication leads to antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and/or complement dependent cytotoxicity (CDC). ADCC and ADCP are mediated through the binding of the Fc region to Fc receptors on the surface of cells of the immune system. CDC is mediated through the binding of the Fc with proteins of the complement system, e.g., C1q. In some embodiments, the bispecific antigen binding proteins of the invention comprise one or more amino acid substitutions in the constant region to enhance effector function, including ADCC activity, CDC activity, ADCP activity, and/or the clearance or half-life of the antigen binding protein. Exemplary amino acid substitutions (EU numbering) that can enhance effector function include, but are not limited to, E233L, L2341, L234Y, L235S, G236A, S239D, F243L, F243V, P2471, D280H, K290S, K290E, K290N, K290Y, R292P, E294L, Y296W, S298A, S298D, S298V, S298G, S298T, T299A, Y300L, V305I, Q311M, K326A, K326E, K326W, A330S, A330L, A330M, A330F, 1332E, D333A, E333S, E333A, K334A, K334V, A339D, A339Q, P396L, or combinations of any of the foregoing. 
     In other embodiments, the bispecific antigen binding proteins of the invention comprise one or more amino acid substitutions in the constant region to reduce effector function. Exemplary amino acid substitutions (EU numbering) that can reduce effector function include, but are not limited to, C220S, C226S, C229S, E233P, L234A, L234V, V234A, L234F, L235A, L235E, G237A, P238S, S267E, H268Q, N297A, N297G, V309L, E318A, L328F, A330S, A331S, P331S or combinations of any of the foregoing. 
     Exemplary substitutions to aid in correct assembly of hetero Ig molecules are shown in  FIG. 1 . Preferred substitutions for the hetero Ig format are shown in (v2) in  FIG. 1  and Table N. All positions in Table N are according to EU numbering. 
     
       
         
           
               
             
               
                 TABLE N 
               
             
            
               
                   
               
               
                 Mutations in the hetero-Ig Molecules 
               
            
           
           
               
               
               
               
            
               
                 Chain 
                 Domain 
                 Mutation 
                 EU # 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Light chain 1 
                 Variable 
                 E 
                 38 
               
               
                   
                 Constant 
                 E 
                 176 
               
               
                 Light chain 2 
                 Variable 
                 K 
                 38 
               
               
                   
                 Constant 
                 K 
                 176 
               
               
                 Heavy chain 1 
                 Variable 
                 K 
                 39 
               
               
                   
                 CH1 
                 K 
                 183 
               
               
                   
                 CH3 
                 D 
                 392 
               
               
                   
                 CH3 
                 D 
                 409 
               
               
                 Heavy chain 2 
                 Variable 
                 E 
                 39 
               
               
                   
                 CH1 
                 E 
                 183 
               
               
                   
                 CH3 
                 K 
                 356 
               
               
                   
                 CH3 
                 K 
                 399 
               
               
                   
               
            
           
         
       
     
     Preferred charged amino acids for the IgG-Fab format, as well as CH/CL domain swaps as described herein for  FIG. 26 , are shown in Table 0. “Fab 1” and “Fab 2” in Table 0 are as depicted in  FIGS. 25 and 26 . Fab 1 is at the N-terminus of the IgG-Fab molecule and Fab 2 is at its C-terminus. The identity and position of the charged amino acids in the Fab appear to the right of the domain listed in Table 0. All positions are according to EU numbering. 
     
       
         
           
               
             
               
                 TABLE O 
               
             
            
               
                   
               
               
                 Mutations in the I2G-Fab Molecules 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 EU  
                   
                   
                 EU  
               
               
                 Designation 
                 Domain 
                 Mutation 
                 # 
                 Domain 
                 Mutation 
                 # 
               
               
                   
               
               
                 CPMv1 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                   
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                 CPMv1 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                 Fab 1 swap 
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                 CPMv1 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 2 swap 
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                 CPMv1 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 1&amp;2 
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                 swap 
                   
                   
                   
                   
                   
                   
               
               
                 CPMv2 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                   
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                  46 
                 Fab 2 VL 
                 E 
                  46 
               
               
                   
                 Fab 1 VH 
                 K 
                  46 
                 Fab 2 VH 
                 K 
                  46 
               
               
                 CPMv2 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                 Fab 1 swap 
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                  46 
                 Fab 2 VL 
                 E 
                  46 
               
               
                   
                 Fab 1 VH 
                 K 
                  46 
                 Fab 2 VH 
                 K 
                  46 
               
               
                 CPMv2 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 2 swap 
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                  46 
                 Fab 2 VL 
                 E 
                  46 
               
               
                   
                 Fab 1 VH 
                 K 
                  46 
                 Fab 2 VH 
                 K 
                  46 
               
               
                 CPMv2 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 1 &amp; 2  
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                 swap 
                 Fab 1 VL 
                 E 
                  46 
                 Fab 2 VL 
                 E 
                  46 
               
               
                   
                 Fab 1 VH 
                 K 
                  46 
                 Fab 2 VH 
                 K 
                  46 
               
               
                 CPMv3 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                   
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                 141 
                 Fab 2 VL 
                 E 
                  51 
               
               
                   
                 Fab 1 VH 
                 K 
                  51 
                 Fab 2 VH 
                 K 
                 141 
               
               
                 CPMv3 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 K 
                 230 
               
               
                 Fab 1 swap 
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 E 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                 141 
                 Fab 2 VL 
                 E 
                  51 
               
               
                   
                 Fab 1 VH 
                 K 
                  51 
                 Fab 2 VH 
                 K 
                 141 
               
               
                 CPMv3 
                 Fab 1 CL 
                 E 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 2 swap 
                 Fab 1 CH1 
                 K 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                   
                 Fab 1 VL 
                 E 
                 141 
                 Fab 2 VL 
                 E 
                  51 
               
               
                   
                 Fab 1 VH 
                 K 
                  51 
                 Fab 2 VH 
                 K 
                 141 
               
               
                 CPMv3 
                 Fab 1 CL 
                 K 
                 230 
                 Fab 2 CL 
                 E 
                 230 
               
               
                 Fab 1 &amp; 2  
                 Fab 1 CH1 
                 E 
                 230 
                 Fab 2 CH1 
                 K 
                 230 
               
               
                 swap 
                 Fab 1 VL 
                 E 
                 141 
                 Fab 2 VL 
                 E 
                  51 
               
               
                   
                 Fab 1 VH 
                 K 
                  51 
                 Fab 2 VH 
                 K 
                 141 
               
               
                   
               
            
           
         
       
     
     Binding Affinity and Biological Activity 
     In another embodiment of the foregoing aspects of the invention, an anti-TL1A antibody (or an antigen-binding fragment thereof) or the TL1A binding entity of a hetero Ig bispecific antigen binding protein or an IgG-scFv antigen binding protein binds TL1A with a binding affinity (K D1 ) of at least 1×10 −10  M −1 , at least 5×10 −10  M −1 , at least 1×10 −10  M −1 , at least 5×10 −10  M −1 , at least 8×10 −10  M −1 , or at least 1×10 −10  M −1  wherein the binding affinity is measured by surface plasmon resonance, such as Biacore. More particularly, the invention relates to the following embodiments: 
     an anti-TL1A antibody (or an antigen-binding fragment thereof) binds to human TL1A when the antibody is immobilized on an SCMS sensor chip at a binding affinity of about 1 to about 5 KD pM or to cynomolgus TL1A when the antibody is immobilized on an SCMS sensor chip at a binding affinity of about 1 −10  to about 7.5 −10  M −1  KD; 
     an anti-TNF-α binding entity of a hetero Ig bispecific antigen binding protein binds to human TNF-α when the antigen binding protein is immobilized on an SCMS sensor chip at a binding affinity of about 1 −10  to about 5 −10  M −1  KD pM, preferably about 10 to about 12 pM; 
     an anti-TL1A binding entity of a hetero Ig bispecific antigen binding protein binds to human TL1A when the antigen binding protein is immobilized on an SCMS sensor chip at a binding affinity of about 2 −10  to about 6.5 −10  M −1  KD, preferably about 10 to about 11 pM; 
     an anti-TNF-α binding entity of an IgG-scFv bispecific antigen binding protein binds to human TNF-α when the antigen binding protein is immobilized on an SCMS sensor chip at a binding affinity of about 4.5 −10  to about 7.5 −10  M −1  KD, preferably about 10 to about 20 pM; and 
     a TL1A binding entity of an IgG-scFv bispecific antigen binding protein binds to human TL1A when the antigen binding protein is immobilized on an SCMS sensor chip at a binding affinity of about 1.5 to about 160 pM KD. 
     Further details on TL1A and TNF-α binding appear in Examples 13 and 15 hereinafter. 
     In another embodiment of the foregoing aspects of the invention, a TNF-α binding entity of an IgG-Fab bispecific antigen binding protein binds a TNF-α trimer with a binding affinity (K D1 ) of 1×10 −10  M −1 , at least 5×10 −10  M −1 , at least 1×10 −10  M −1 , at least 5×10 −10  M −1 , at least 8×10 −10  M −1 , or at least 1×10 −10  M −1  wherein the binding affinity is measured by surface plasmon resonance, such as Biacore. 
     In other embodiments of the foregoing aspects of the invention: 
     an anti-TL1A antibody (or an antigen-binding fragment thereof) neutralizes or inhibits human TL1A in a TF-1 NF-KB reporter cell line with an IC 50  of about 0.08 to about 3 nM, cynomolgus TL1A TL1A in a TF-1 NF-κB reporter cell line with an IC 50  of about 0.375 to about 10 nM; 
     a TNF-α binding entity of a hetero Ig bispecific antigen binding protein neutralizes or inhibits human TNF-α in a TF-1 NF-KB reporter cell line with an IC 50  of about 50 to 550 pM, with about 50 to about 110 pM preferred; 
     a TL1A binding entity of a hetero Ig bispecific antigen binding protein neutralizes or inhibits human TL1A in a TF-1 NF-KB reporter cell line with an IC 50  of about 45 to about 3500 pM, with about 45 to 190 pM preferred; 
     a TNF-α binding entity of an IgG-scFv bispecific antigen binding protein neutralizes or inhibits soluble human TNF-α in a TF-1 NF-KB reporter cell line with an IC 50  of about 20 to about 75 pM; and 
     a TL1A binding entity of an IgG-scFv bispecific antigen binding protein neutralizes or inhibits human TL1A in a TF-1 NF-KB reporter cell line with an IC 50  of about 19 to about 200 pM. 
     Further details on TL1A inhibition and TNF-α inhibition appears in Examples 14 and 16 hereinafter. 
     Immunoconjugates, Derivatives, Variants 
     The antibodies and bispecific antibodies of the invention may be used alone or as immunoconjugates with a therapeutic agent (e.g., a cytotoxic agent). In some embodiments, the agent is a chemotherapeutic agent. In some embodiments, the agent is a radioisotope, including but not limited to Lead-212, Bismuth-212, Astatine-211, Iodine-131, Scandium-47, Rhenium-186, Rhenium-188, Yttrium-90, Iodine-123, Iodine-125, Bromine-77, Indium-111, and fissionable nuclides such as Boron-10 or an Actinide. In other embodiments, the agent is a toxin or cytotoxic drug, including but not limited to ricin, abrin, modified Pseudomonas enterotoxin A, Pseudomonas exotoxin, calicheamicin, adriamycin, 5-fluorouracil, diphtheria toxin, and the like. Methods of conjugation of antibodies to such agents are known in the literature, and include direct and indirect conjugation. 
     Suitable detectable molecules may be directly or indirectly attached to the antibodies and bispecific antibodies of the present invention. Suitable detectable molecules include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent markers, chemiluminescent markers, magnetic particles and the like. For indirect attachment of a detectable or cytotoxic molecule, the detectable or cytotoxic molecule can be conjugated with a member of a complementary/anti-complementary pair, where the other member is bound to the binding polypeptide or antibody portion. For these purposes, biotin/streptavidin is an exemplary complementary/anti-complementary pair. 
     The bispecific antibodies and antibodies of the invention also include derivatives that are modified, e.g., by the covalent attachment of any type of molecule to the antibody such that covalent attachment does not prevent the antibody from binding to its epitope. Examples of suitable derivatives include but are not limited to antibodies or bispecific antibodies that are fucosylated, glycosylated, acetylated, pegylated, phosphorylated, or amidated. The antibodies and bispecific antibodies and derivatives thereof of the invention may themselves by derivatized by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other proteins, and the like. In some embodiments of the invention, at least one heavy chain of the antibody or bispecific antigen binding protein is PEGylated. In some embodiments, the PEGylation is N-linked or is linked through the sidechain of an amino acid (e.g., lysine). 
     Glycosylation can contribute to the effector function of antibodies, particularly IgG1 antibodies. Thus, in some embodiments, the antigen binding proteins of the invention may comprise one or more amino acid substitutions that affect the level or type of glycosylation of the binding proteins. Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tri-peptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tri-peptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used. 
     In certain embodiments, glycosylation of the antigen binding proteins described herein is increased by adding one or more glycosylation sites, e.g., to the Fc region of the binding protein. Addition of glycosylation sites to the antigen binding protein can be conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tri-peptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the starting sequence (for O-linked glycosylation sites). For ease, the antigen binding protein amino acid sequence may be altered through changes at the DNA level, particularly by mutating the DNA encoding the target polypeptide at preselected bases such that codons are generated that will translate into the desired amino acids. 
     The invention also encompasses production of antigen binding proteins with altered carbohydrate structure resulting in altered effector activity, including antigen binding proteins with absent or reduced fucosylation that exhibit improved ADCC activity. Various methods are known in the art to reduce or eliminate fucosylation. For example, ADCC effector activity is mediated by binding of the antibody molecule to the FcγRIII receptor, which has been shown to be dependent on the carbohydrate structure of the N-linked glycosylation at the N297 residue of the CH2 domain. Non-fucosylated antibodies bind this receptor with increased affinity and trigger FcγRIII-mediated effector functions more efficiently than native, fucosylated antibodies. For example, recombinant production of non-fucosylated antibody in CHO cells in which the alpha-1,6-fucosyl transferase enzyme has been knocked out results in antibody with 100-fold increased ADCC activity (see Yamane-Ohnuki et al., Biotechnol Bioeng. 87(5):614-22, 2004). Similar effects can be accomplished through decreasing the activity of alpha-1,6-fucosyl transferase enzyme or other enzymes in the fucosylation pathway, e.g., through siRNA or antisense RNA treatment, engineering cell lines to knockout the enzyme(s), or culturing with selective glycosylation inhibitors (see Rothman et al., Mol Immunol. 26(12):1113-23, 1989). Some host cell strains, e.g. Lec13 or rat hybridoma YB2/0 cell line naturally produce antibodies with lower fucosylation levels (see Shields et al., J Biol Chem. 277(30):26733-40, 2002 and Shinkawa et al., J Biol Chem. 278(5):3466-73, 2003). An increase in the level of bisected carbohydrate, e.g. through recombinantly producing antibody in cells that overexpress GnTIII enzyme, has also been determined to increase ADCC activity (see Umana et al., Nat Biotechnol. 17(2):176-80, 1999). 
     In other embodiments, glycosylation of the antigen binding proteins described herein is decreased or eliminated by removing one or more glycosylation sites, e.g., from the Fc region of the binding protein. Amino acid substitutions that eliminate or alter N-linked glycosylation sites can reduce or eliminate N-linked glycosylation of the antigen binding protein. In certain embodiments, the bispecific antigen binding proteins described herein comprise a mutation at position N297 (EU numbering), such as N297Q, N297A, or N297G. In one particular embodiment, the bispecific antigen binding proteins of the invention comprise a Fc region from a human IgG1 antibody with a N297G mutation. To improve the stability of molecules comprising a N297 mutation, the Fc region of the molecules may be further engineered. For instance, in some embodiments, one or more amino acids in the Fc region are substituted with cysteine to promote disulfide bond formation in the dimeric state. Residues corresponding to V259, A287, R292, V302, L306, V323, or I332 (EU numbering) of an IgG1 Fc region may thus be substituted with cysteine. In one embodiment, specific pairs of residues are substituted with cysteine such that they preferentially form a disulfide bond with each other, thus limiting or preventing disulfide bond scrambling. In certain embodiments pairs include, but are not limited to, A287C and L306C, V259C and L306C, R292C and V302C, and V323C and I332C. In particular embodiments, the bispecific antigen binding proteins described herein comprise a Fc region from a human IgG1 antibody with mutations at R292C and V302C. In such embodiments, the Fc region may also comprise a N297G mutation. 
     Modifications of the antigen binding proteins of the invention to increase serum half-life also may desirable, for example, by incorporation of or addition of a salvage receptor binding epitope (e.g., by mutation of the appropriate region or by incorporating the epitope into a peptide tag that is then fused to the antigen binding protein at either end or in the middle, e.g., by DNA or peptide synthesis; see, e.g., WO96/32478) or adding molecules such as PEG or other water soluble polymers, including polysaccharide polymers. The salvage receptor binding epitope preferably constitutes a region wherein any one or more amino acid residues from one or two loops of a Fc region are transferred to an analogous position in the antigen binding protein. In one embodiment, three or more residues from one or two loops of the Fc region are transferred. In one embodiment, the epitope is taken from the CH2 domain of the Fc region (e.g., an IgG Fc region) and transferred to the CH1, CH3, or VH region, or more than one such region, of the antigen binding protein. Alternatively, the epitope is taken from the CH2 domain of the Fc region and transferred to the CL region or VL region, or both, of the antigen binding protein. See International applications WO 97/34631 and WO 96/32478 for a description of Fc variants and their interaction with the salvage receptor. 
     The bispecific antibodies and antibodies of the invention include variants having single or multiple amino acid substitutions, deletions, additions, or replacements that retain their biological properties (e.g., blocking the binding of TL1A and/or TNF-α to their respective receptors, inhibiting the biological activity of TL1A and TNF-α). A person of ordinary skill in the art can produce variants having single or multiple amino acid substitutions, deletions, additions or replacements. These variants may include, inter alia: (a) variants in which one or more amino acid residues are substituted with conservative or non-conservative amino acids, (b) variants in which one or more amino acids are added to or deleted from the polypeptide, (c) variants in which one or more amino acids include a substituent group, and (d) variants in which the polypeptide is fused with another peptide or polypeptide such as a fusion partner, a protein tag or other chemical moiety, that may confer useful properties to the polypeptide, such as, for example, an epitope for an antibody, a polyhistidine sequence, a biotin moiety and the like. Antibodies and bispecific antibodies of the invention may include variants in which amino acid residues from one species are substituted for the corresponding residue in another species, either at the conserved or non-conserved positions. In another embodiment, amino acid residues at non-conserved positions are substituted with conservative or non-conservative residues. The techniques for obtaining these variants, including genetic (suppressions, deletions, mutations, etc.), chemical, and enzymatic techniques, are known to the person having ordinary skill in the art. 
     Nucleic Acids, Vectors, Host Cells 
     The invention also includes isolated nucleic acids encoding the bispecific antibodies of the invention, which includes, for instance, the light chain, light chain variable region, light chain constant region, heavy chain, heavy chain variable region, heavy chain constant region, linkers, and any and all components and combinations thereof of the bispecific antibodies disclosed herein. Nucleic acids of the invention include nucleic acids having at least 80%, more preferably at least about 90%, more preferably at least about 95%, and most preferably at least about 98% homology to nucleic acids of the invention. The terms “percent similarity”, “percent identity” and “percent homology” when referring to a particular sequence are used as set forth in the University of Wisconsin GCG® software program. Nucleic acids of the invention also include complementary nucleic acids. In some instances, the sequences will be fully complementary (no mismatches) when aligned. In other instances, there may be up to about a 20% mismatch in the sequences. In some embodiments of the invention are provided nucleic acids encoding both a heavy chain and a light chain of an antibody of the invention. 
     Nucleic acids of the invention can be cloned into a vector, such as a plasmid, cosmid, bacmid, phage, artificial chromosome (BAC, YAC) or virus, into which another genetic sequence or element (either DNA or RNA) may be inserted so as to bring about the replication of the attached sequence or element. In some embodiments, the expression vector contains a constitutively active promoter segment (such as but not limited to CMV, SV40, Elongation Factor or LTR sequences) or an inducible promoter sequence such as the steroid inducible pIND vector (Invitrogen), where the expression of the nucleic acid can be regulated. Expression vectors of the invention may further comprise regulatory sequences, for example, an internal ribosomal entry site. The expression vector can be introduced into a cell by transfection, for example. 
     For IgG-Fab bispecific antigen binding proteins, nucleic acids encoding each of the three components may be cloned into the same expression vector. In some embodiments, the nucleic acid encoding the light chain of the IgG-Fab molecule and the nucleic acid encoding the second polypeptide (which comprises the other half of the C-terminal Fab domain) are cloned into one expression vector, whereas the nucleic acid encoding the modified heavy chain (fusion protein comprising a heavy chain and half of a Fab domain) is cloned into a second expression vector. In certain embodiments, all components of the bispecific antigen binding proteins described herein are expressed from the same host cell population. For example, even if one or more components is cloned into a separate expression vector, the host cell is co-transfected with both expression vectors such that one cell produces all components of the bispecific antigen binding proteins. 
     In another embodiment, the present invention provides an expression vector comprising the following operably linked elements; a transcription promoter; a first nucleic acid molecule encoding the heavy chain of a bispecific antigen binding protein, antibody or antigen-binding fragment of the invention; a second nucleic acid molecule encoding the light chain of a bispecific antigen binding protein, antibody or antigen-binding fragment of the invention; and a transcription terminator. In another embodiment, the present invention provides an expression vector comprising the following operably linked elements; a first transcription promoter; a first nucleic acid molecule encoding the heavy chain of a bispecific antigen binding protein, antibody or antigen-binding fragment of the invention; a first transcription terminator; a second transcription promoter a second nucleic acid molecule encoding the light chain of a bispecific antigen binding protein, antibody or antigen-binding fragment of the invention; and a second transcription terminator. 
     A secretory signal peptide sequence can also, optionally, be encoded by the expression vector, operably linked to the coding sequence of interest, so that the expressed polypeptide can be secreted by the recombinant host cell, for more facile isolation of the polypeptide of interest from the cell, if desired. For instance, in some embodiments, signal peptide sequences may be appended/fused to the amino terminus of any of the polypeptide sequences listed in Tables E, J, K, and L. In certain embodiments, a signal peptide having the amino acid sequence of MKHLWFFLLLVAAPRWVLS (SEQ ID NO: 499) is fused to the amino terminus of any of the polypeptide sequences in Tables D, I, J, and K. In other embodiments, a signal peptide having the amino acid sequence of METPAQLLFLLLLWLPDTTG (SEQ ID NO: 501) is fused to the amino terminus of any of the polypeptide sequences in Tables E, J, K, and L. In still other embodiments, a signal peptide having the amino acid sequence of MDMRVPAQLLGLLLLWLRGARC (SEQ ID NO: 503) is fused to the amino terminus of any of the polypeptide sequences in Tables E, J, K, and L. Each of the foregoing signal peptides is encoded by the nucleic acid having a sequence immediately preceding it in the Sequence Listing. Other suitable signal peptide sequences that can be fused to the amino terminus of the polypeptide sequences described herein include: MEAPAQLLFLLLLWLPDTTG (SEQ ID NO: 504), MEWTWRVLFLVAAATGAHS (SEQ ID NO: 505), and MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 506). Other signal peptides are known to those of skill in the art and may be fused to any of the polypeptide chains listed in Tables E, J, K and L, for example, to facilitate or optimize expression in particular host cells. 
     Recombinant host cells comprising such vectors and expressing the heavy and light chains are also provided. 
     Antibody-producing cells and bispecific antigen binding protein producing cells contain, depending on the bispecific antigen binding protein format, nucleic acids encoding the heavy chain, light chain, heavy chain-scFv construct, heavy chain-Fab heavy chain variable domain construct, and Fab light chain variable domain. Such nucleic acids can be used to produce the antibodies or bispecific antibodies of the invention in accordance with techniques known in the art. The present invention, in one embodiment, provides a method of producing a bispecific antigen binding protein or antibody comprising culturing a recombinant host cell expressing the heavy and light chains or other constructs noted above and isolating the bispecific antigen binding protein or antibody produced by the cell. 
     The recombinant host cell may be a prokaryotic cell, for example an  E. coli  cell, or a eukaryotic cell, for example a mammalian cell or a yeast cell. Yeast cells include  Saccharomyces cerevisiae, Schizosaccharomyces pombe,  and  Pichia pastoris  cells. Mammalian cells include VERO, HeLa, Chinese hamster Ovary (CHO), W138, baby hamster kidney (BHK), COS-7, MDCK, human embryonic kidney line 293, normal dog kidney cell lines, normal cat kidney cell lines, monkey kidney cells, African green monkey kidney cells, COS cells, and non-tumorigenic mouse myoblast G8 cells, fibroblast cell lines, myeloma cell lines, mouse NIH/3T3 cells, LMTK31 cells, mouse sertoli cells, human cervical carcinoma cells, buffalo rat liver cells, human lung cells, human liver cells, mouse mammary tumor cells, TRI cells, MRC 5 cells, and FS4 cells. Antibody-producing and bispecific antigen binding protein-producing cells of the invention also include any insect expression cell line known, such as for example,  Spodoptera frugiperda  cells. In a preferred embodiment, the cells are mammalian cells. In a most preferred embodiment, the mammalian cells are CHO cells. 
     The antibody-producing cells preferably are substantially free of TL1A and TNF-α binding competitors. In preferred embodiments, the antibody-producing cells comprise less than about 10%, preferably less than about 5%, more preferably less than about 1%, more preferably less than about 0.5%, more preferably less than about 0.1%, and most preferably 0% by weight TL1A or TNF-α binding competitors. In some embodiments, the antibodies and bispecific antibodies produced are substantially free of TL1A and TNF-α competitors. In preferred embodiments, the antibodies and bispecific antibodies produced comprise less than about 10%, preferably less than about 5%, more preferably less than about 1%, more preferably less than about 0.5%, more preferably less than about 0.1%, and most preferably 0% by weight both TL1A and TNF-α binding competitors. 
     Purification 
     Methods of antibody purification are known in the art and can be employed with production of the antibodies and bispecific antibodies of the present invention. In some embodiments of the invention, methods for antibody purification include filtration, affinity column chromatography, cation exchange chromatography, anion exchange chromatography, and concentration. The filtration step preferably comprises ultrafiltration, and more preferably ultrafiltration and diafiltration. Filtration is preferably performed at least about 5-50 times, more preferably 10 to 30 times, and most preferably 14 to 27 times. Affinity column chromatography, may be performed using, for example, PROSEP® Affinity Chromatography (Millipore, Billerica, Mass.). In a preferred embodiment, the affinity chromatography step comprises PROSEP®-vA column chromatography. Eluate may be washed in a solvent detergent. Cation exchange chromatography may include, for example, SP-Sepharose Cation Exchange Chromatography. Anion exchange chromatography may include, for example but not limited to, Q-Sepharose Fast Flow Anion Exchange. The anion exchange step is preferably non-binding, thereby allowing removal of contaminants including DNA and BSA. The antibody product is preferably nanofiltered, for example, using a Pall DV 20 Nanofilter. The antibody product may be concentrated, for example, using ultrafiltration and diafiltration. The method may further comprise a step of size exclusion chromatography to remove aggregates. Further parameters of purification appear in the working examples hereinafter. 
     The bispecific antibodies, antibodies or antigen-binding fragments may also be produced by other methods known in the art, for example by chemical coupling of antibodies and antibody fragments. 
     Uses of the Monospecific and Bispecific Antibodies of the Invention 
     The bispecific antibodies and monospecific antibodies of the present invention are useful, for example, for the inhibition of proinflammatory cytokines, such as TL1A and TNF-α. The bispecific antibodies and monospecific antibodies of the invention can be used to treat inflammatory disorders and autoimmune diseases, such as inflammatory bowel disease (IBD), Crohn&#39;s disease (CD), ulcerative colitis (UC), irritable bowel syndrome (IBS), bladder syndrome/intersticial cystitis, urinary bowel disfunction, sepsis, uveitis, encephalomyelitis, myasthenia gravis, Sjogren&#39;s syndrome (SS), scleroderma, multiple sclerosis (MS), cystic fibrosis (CF), inflammation in chronic kidney disease (CKD), psoriasis (Pso), psoriatic arthritis (PsA), ankylosing spondylitis (AS), rheumatoid arthritis (RA), juvenile rheumatoid arthritis (JRA), osteoarthritis (OA), spondyloarthropathy, primary sclerosing cholangitis, primary biliary cirrhosis, atherosclerosis, splenomegaly, inflammation in chronic kidney disease (CKD), atopic dermatitis (AD), eczematous dermatitis, contact dermatitis systemic sclerosis, systemic lupus erythematosus (SLE), lupus nephritis (LN), cutaneous lupus erythematosus, autoimmune thyroiditis, IgA nephropathy, diabetic kidney disease, antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV), minimal change disease (lipoid nephrosis), focal segmental glomerulosclerosis (FSGS), nephrogenic systemic fibrosis (NSF), nephrogenic fibrosing dermopathy, fibrosing cholestatic hepatitis, eosinophilic fasciitis (Shulman&#39;s syndrome), scleromyxedema (popular mucinosis), scleroderma, lichen sclerosusetatrophicus, inflammatory lung injury such as idiopathic pulmonary fibrosis, asthma, chronic obstructive pulmonary disease (COPD), airway hyper-responsiveness, chronic bronchitis, allergic asthma, eczema,  Helicobacter pylori  infection, intraabdominal adhesions and/or abscesses as results of peritoneal inflammation (e.g., from infection, injury, etc.), nephrotic syndrome, idiopathic demyelinating polyneuropathy, Guillain-Barre syndrome, transplant rejection, organ allograft rejection, graft vs. host disease (GVHD) (e.g., from a transplant, such as blood, bone marrow, kidney, pancreas, liver, orthotopic liver, lung, heart, intestine, small intestine, large intestine, thymus, allogeneic stem cell, reduced-intensity allogeneic, bone, tendon, cornea, skin, heart valves, veins, arteries, blood vessels, stomach and testis), IgA nephropathy, diabetic kidney disease, diabetes mellitus, minimal change disease (lipoid nephrosis), nephrogenic systemic fibrosis (NSF), nephrogenic fibrosing dermopathy, fibrosing cholestatic hepatitis, eosinophilic fasciitis (Shulman&#39;s syndrome), scleromyxedema (popular mucinosis), scleroderma, lichen sclerosusetatrophicus, Takatsuki disease (or PEP syndrome), nephrotic syndrome, POEMs syndrome, Crow-Fukase syndrome, nephrotic syndrome, antineutrophil cytoplasmic antibodies, vasculitis, giant cell arteritis and multiple-myeloma-induced lytic bone disease, streptococcal cell wall (SCW)-induced arthritis, gingivitis/periodontitis, herpetic stromal keratitis, gluten-sensitive enteropathy restenosis, Kawasaki&#39;s disease, and immune-mediated renal diseases. The bispecific antibodies and monospecific antibodies described herein can also be used to treat cancer, including angiogenesis. 
     In one embodiment, the invention concerns methods of treating one or more of the aforementioned diseases and disorders in a mammal in need of such treatment by administering a therapeutically effective amount of a monospecific or bispecific antigen binding protein of the present invention. In a preferred embodiment the mammal is a human. In another preferred embodiment, the disease or disorder is IBD, CD, or UC. 
     The invention further concerns the use of the bispecific and monospecific antibodies of the present invention in the treatment of inflammatory diseases characterized by the presence of elevated levels of TL1A or/(in the case of the bispecific antibodies) TNF-α, and in the treatment of cancers characterized by the presence of elevated levels of TL1A and/or TNF-α. 
     Accordingly, in one embodiment, the present invention provides a method of inhibiting one or more of proinflammatory cytokines, e.g., TL1A and TNF-α, in a mammal in need of such treatment comprising administering a therapeutically effective amount of a bispecific or monospecific antibody of the invention to a mammal in need of such treatment. In a preferred embodiment, the mammal is a human. The method may be used to treat a disorder characterized by elevated expression or activity of TL1A or TNF-α. The monospecific or bispecific antigen binding protein may be administered with another pharmaceutical agent, either in the same formulation or separately. 
     In another embodiment, the present invention provides a composition comprising a monospecific or bispecific antigen binding protein as described herein and a pharmaceutically acceptable carrier. A pharmaceutical composition comprising an antibody, e.g., a bispecific antigen binding protein, of the invention can be formulated according to known methods to prepare pharmaceutically useful compositions, whereby the therapeutic antibodies are combined in a mixture with a pharmaceutically acceptable carrier. A composition is said to comprise a “pharmaceutically acceptable carrier” if its administration can be tolerated by a recipient patient. Sterile phosphate-buffered saline is one example of a pharmaceutically acceptable carrier. Other suitable carriers are well-known to those in the art. See, for example, Getman), ed., Remington&#39;s Pharmaceutical Sciences, 19th Edition, Mack Publishing Company (1995). 
     For pharmaceutical use, an antibody, e.g., a bispecific antigen binding protein, of the present invention is formulated for parenteral, particularly intravenous or subcutaneous, delivery according to conventional methods. Intravenous administration may be by bolus injection, controlled release, e.g., using mini-pumps or other appropriate technology, or by infusion over a typical period of one to several hours. In general, pharmaceutical formulations will include an antibody, e.g., a bispecific antigen binding protein, of the invention in combination with a pharmaceutically acceptable carrier, such as saline, buffered saline, 5% dextrose in water or the like. Formulations may further include one or more excipients, preservatives, solubilizers, buffering agents, albumin to prevent protein loss on vial surfaces, etc. When utilizing such a combination therapy, the antibodies, which include bispecific antibodies, may be combined in a single formulation or may be administered in separate formulations. Methods of formulation are well known in the art and are disclosed, for example, in Gennaro, ed., Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton Pa. (1990), which is incorporated herein by reference. Therapeutic doses will generally be in the range of 0.1 to 100 mg/kg of patient weight per day, preferably 0.5-20 mg/kg per day, with the exact dose determined by the clinician according to accepted standards, taking into account the nature and severity of the condition to be treated, patient traits, etc. Determination of dose is within the level of ordinary skill in the art. More commonly, the antibodies will be administered over one week or less, often over a period of one to three days. Generally, the dosage of administered antibodies will vary depending upon such factors as the patient&#39;s age, weight, height, sex, general medical condition and previous medical history. Typically, it is desirable to provide the recipient with a dosage of antibodies which is in the range of from about 1 pg/kg to 10 mg/kg (amount of agent/body weight of patient), although a lower or higher dosage also may be administered as circumstances dictate. 
     Administration of an antibody, e.g., bispecific antigen binding protein, of the invention to a subject can be intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, intrapleural, intrathecal, by perfusion through a regional catheter, or by direct intralesional injection. When administering therapeutic antibodies by injection, the administration may be by continuous infusion or by single or multiple boluses. 
     Additional routes of administration include oral, mucosal-membrane, pulmonary, and transcutaneous. Oral delivery is suitable for polyester microspheres, zein microspheres, proteinoid microspheres, polycyanoacrylate microspheres, and lipid-based systems (see, for example, DiBase et al., “Oral Delivery of Microencapsulated Proteins”, in Sanders et al., eds., Protein Delivery: Physical Systems, pp. 255-288, Plenum Press (1997)). The feasibility of an intranasal delivery is exemplified by such a mode of insulin administration (see, for example, Hinchcliffe et al., Adv. Drug Deliv. Rev., 35:199 (1999)). Dry or liquid particles comprising antibodies of the invention can be prepared and inhaled with the aid of dry-powder dispersers, liquid aerosol generators, or nebulizers (e.g., Pettit et al., TIBTECH, 16:343 (1998); Patton et al., Adv. Drug Deliv. Rev., 35:235 (1999)). This approach is illustrated by the AERX® diabetes management system, which is a hand-held electronic inhaler that delivers aerosolized insulin into the lungs. Studies have shown that proteins as large as 48,000 kDa have been delivered across skin at therapeutic concentrations with the aid of low-frequency ultrasound, which illustrates the feasibility of transcutaneous administration (Mitragotri et al., Science, 269:850 (1995)). Transdermal delivery using electroporation provides another means to administer a molecule having IL-17 and TNF-α/p19 binding activity (Potts et al., Pharm. Biotechnol., 10:213 (1997)). 
     For purposes of therapy, compositions comprising an antibody, e.g., a bispecific antigen binding protein, of the invention and a pharmaceutically acceptable carrier are administered to a patient in a therapeutically effective amount. A combination of an antibody, e.g., a bispecific antigen binding protein, of the present invention and a pharmaceutically acceptable carrier is said to be administered in a “therapeutically effective amount” if the amount administered is physiologically significant. An agent is physiologically significant if its presence results in a detectable change in the physiology of a recipient patient. For example, an agent used to treat inflammation is physiologically significant if its presence alleviates the inflammatory response. Effective treatment may be assessed in a variety of ways. In one embodiment, effective treatment is determined by reduced inflammation. In other embodiments, effective treatment is marked by inhibition of inflammation. In still other embodiments, effective therapy is measured by increased well-being of the patient including such signs as weight gain, regained strength, decreased pain, thriving, and subjective indications from the patient of better health. 
     A pharmaceutical composition comprising an antibody, e.g., a bispecific antigen binding protein, of the invention can be furnished in liquid form, in an aerosol, or in solid form. Liquid forms, are illustrated by injectable solutions and oral suspensions. Exemplary solid forms include capsules, tablets, and controlled-release forms. The latter form is illustrated by miniosmotic pumps and implants (Bremer et al., Pharm. Biotechnol., 10:239 (1997); Ranade, “Implants in Drug Delivery”, in Ranade et al., eds., Drug Delivery Systems, pp. 95-123, CRC Press (1995); Bremer et al., “Protein Delivery with Infusion Pumps”, in Sanders et al., eds., Protein Delivery: Physical Systems, pp. 239-254, Plenum Press (1997); Yewey et al., “Delivery of Proteins from a Controlled Release Injectable Implant”, in Sanders et al., eds., Protein Delivery: Physical Systems, pp. 93-117, Plenum Press (1997). 
     Liposomes provide one means to deliver therapeutic polypeptides to a subject intravenously, intraperitoneally, intrathecally, intramuscularly, subcutaneously, or via oral administration, inhalation, or intranasal administration. Liposomes are microscopic vesicles that consist of one or more lipid bilayers surrounding aqueous compartments (see, generally, Bakker-Woudenberg et al., Eur. J. Clin. Microbiol. Infect. Dis., 12(Suppl. 1):561 (1993), Kim, Drugs, 46:618 (1993), and Ranade, “Site-Specific Drug Delivery Using Liposomes as Carriers”, in Ranade et al., eds., Drug Delivery Systems, pp. 3-24, CRC Press (1995)). Liposomes are similar in composition to cellular membranes and as a result, liposomes can be administered safely and are biodegradable. Depending on the method of preparation, liposomes may be unilamellar or multilamellar, and liposomes can vary in size with diameters ranging from 0.02 .mu.m to greater than 10 .mu.m. A variety of agents can be encapsulated in liposomes: hydrophobic agents partition in the bilayers and hydrophilic agents partition within the inner aqueous space(s) (see, for example, Machy et al., Liposomes in Cell Biology and Pharmacology, John Libbey (1987), and Ostro et al., American J. Hosp. Pharm., 46:1576 (1989)). Moreover, it is possible to control the therapeutic availability of the encapsulated agent by varying liposome size, the number of bilayers, lipid composition, as well as the charge and surface characteristics of the liposomes. 
     Liposomes can absorb to virtually any type of cell and then slowly release the encapsulated agent. Alternatively, an absorbed liposome may be endocytosed by cells that are phagocytic. Endocytosis is followed by intralysosomal degradation of liposomal lipids and release of the encapsulated agents (Scherphof et al., Ann. N. Y. Acad. Sci., 446:368 (1985)). After intravenous administration, small liposomes (0.1 to 1.0 .mu.m) are typically taken up by cells of the reticuloendothelial system, located principally in the liver and spleen, whereas liposomes larger than 3.0 .mu.m are deposited in the lung. This preferential uptake of smaller liposomes by the cells of the reticuloendothelial system has been used to deliver chemotherapeutic agents to macrophages and to tumors of the liver. 
     The reticuloendothelial system can be circumvented by several methods including saturation with large doses of liposome particles, or selective macrophage inactivation by pharmacological means (Claassen et al., Biochim. Biophys. Acta, 802:428 (1984)). In addition, incorporation of glycolipid- or polyethelene glycol-derivatized phospholipids into liposome membranes has been shown to result in a significantly reduced uptake by the reticuloendothelial system (Allen et al.,  Biochim. Biophys. Acta,  1068:133 (1991); Allen et al.,  Biochim. Biophys. Acta,  1150:9 (1993)). 
     Liposomes can also be prepared to target particular cells or organs by varying phospholipid composition or by inserting receptors or ligands into the liposomes. For example, liposomes, prepared with a high content of a nonionic surfactant, have been used to target the liver (Hayakawa et al., Japanese Patent No. 04-244,018; Kato et al., Biol. Pharm. Bull., 16:960 (1993)). These formulations were prepared by mixing soybean phospatidylcholine, .alpha.-tocopherol, and ethoxylated hydrogenated castor oil (HCO-60) in methanol, concentrating the mixture under vacuum, and then reconstituting the mixture with water. A liposomal formulation of dipalmitoylphosphatidylcholine (DPPC) with a soybean-derived sterylglucoside mixture (SG) and cholesterol (Ch) has also been shown to target the liver (Shimizu et al., Biol. Pharm. Bull., 20:881 (1997)). 
     Alternatively, various targeting ligands can be bound to the surface of the liposome, such as antibodies, antibody fragments, carbohydrates, vitamins, and transport proteins. For example, liposomes can be modified with branched type galactosyllipid derivatives to target asialoglycoprotein (galactose) receptors, which are exclusively expressed on the surface of liver cells (Kato et al.,  Crit. Rev. Ther. Drug Carrier Syst.,  14:287 (1997); Murahashi et al.,  Biol. Pharm. Bull.,  20:259 (1997)). Similarly, Wu et al.,  Hepatology,  27:772 (1998), have shown that labeling liposomes with asialofetuin led to a shortened liposome plasma half-life and greatly enhanced uptake of asialofetuin-labeled liposome by hepatocytes. On the other hand, hepatic accumulation of liposomes comprising branched type galactosyllipid derivatives can be inhibited by preinjection of asialofetuin (Murahashi et al.,  Biol. Pharm. Bull.,  20:259 (1997)). Polyaconitylated human serum albumin liposomes provide another approach for targeting liposomes to liver cells (Kamps et al.,  Proc. Nat&#39;l Acad. Sci. USA,  94:11681 (1997)). Moreover, Geho et al. U.S. Pat. No. 4,603,044, describe a hepatocyte-directed liposome vesicle delivery system, which has specificity for hepatobiliary receptors associated with the specialized metabolic cells of the liver. 
     In a more general approach to tissue targeting, target cells are prelabeled with biotinylated antibodies specific for a ligand expressed by the target cell (Harasym et al.,  Adv. Drug Deliv. Rev.,  32: 99 (1998)). After plasma elimination of free antibody, streptavidin-conjugated liposomes are administered. In another approach, targeting antibodies are directly attached to liposomes (Harasym et al.,  Adv. Drug Deliv. Rev.,  32: 99 (1998)). 
     Antibodies can be encapsulated within liposomes using standard techniques of protein microencapsulation (see, for example, Anderson et al.,  Infect. Immun.,  31:1099 (1981), Anderson et al.,  Cancer Res.,  50:1853 (1990), and Cohen et al.,  Biochim. Biophys. Acta,  1063:95 (1991), Alving et al. “Preparation and Use of Liposomes in Immunological Studies”, in Gregoriadis, ed.,  Liposome Technology,  2nd Edition, Vol. III, p. 317, CRC Press (1993), Wassef et al.,  Meth. Enzymol.,  149:124 (1987)). As noted above, therapeutically useful liposomes may contain a variety of components. For example, liposomes may comprise lipid derivatives of poly(ethylene glycol) (Allen et al.,  Biochim. Biophys. Acta,  1150:9 (1993)). 
     Degradable polymer microspheres have been designed to maintain high systemic levels of therapeutic proteins. Microspheres are prepared from degradable polymers such as poly(lactide-co-glycolide) (PLG), polyanhydrides, poly (ortho esters), nonbiodegradable ethylvinyl acetate polymers, in which proteins are entrapped in the polymer (Gombotz et al.,  Bioconjugate Chem.,  6:332 (1995); Ranade, “Role of Polymers in Drug Delivery”, in Ranade et al., eds.,  Drug Delivery Systems, pp.  51-93, CRC Press (1995); Roskos et al., “Degradable Controlled Release Systems Useful for Protein Delivery”, in Sanders et al., eds.,  Protein Delivery: Physical Systems, pp.  45-92, Plenum Press (1997); Bartus et al.,  Science,  281:1161 (1998); Putney et al.,  Nature Biotechnology,  16:153 (1998); Putney,  Curr. Opin. Chem. Biol.,  2:548 (1998)). Polyethylene glycol (PEG)-coated nanospheres can also provide carriers for intravenous administration of therapeutic proteins (see, for example, Gref et al.,  Pharm. Biotechnol.,  10:167 (1996). 
     The formulation can also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Alternatively, or in addition, the composition can comprise an agent that enhances its function, such as, for example, a cytotoxic agent, cytokine, chemotherapeutic agent, or growth-inhibitory agent. Such molecules are suitably present in combination in amounts that are effective for the purpose intended. 
     In one embodiment, an antibody, e.g., a bispecific antigen binding protein, of the invention is administered in combination therapy, i.e., combined with other agents, e.g., therapeutic agents, that are useful for treating pathological conditions or disorders, such as autoimmune disorders and inflammatory diseases. The term “in combination” in this context means that the agents are given substantially contemporaneously, either simultaneously or sequentially. If given sequentially, at the onset of administration of the second compound, the first of the two compounds is preferably still detectable at effective concentrations at the site of treatment. 
     For example, the combination therapy can include one or more an antibodies, e.g., bispecific antibodies, of the invention coformulated with, and/or coadministered with, one or more additional therapeutic agents, e.g., one or more cytokine and growth factor inhibitors, immunosuppressants, anti-inflammatory agents, metabolic inhibitors, enzyme inhibitors, and/or cytotoxic or cytostatic agents, as described in more detail below. Furthermore, one or more antibodies, e.g., bispecific antibodies, described herein may be used in combination with two or more of the therapeutic agents described herein. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the various monotherapies. 
     Preferred therapeutic agents used in combination with an antibody, e.g., bispecific antigen binding protein, of the invention are those agents that interfere at different stages in an inflammatory response. In one embodiment, one or more antibodies, e.g., bispecific antibodies, described herein may be co-formulated with, and/or co-administered with, one or more additional agents such as other cytokine or growth factor antagonists (e.g., soluble receptors, peptide inhibitors, small molecules, ligand fusions); or antibodies or antigen binding fragments thereof that bind to other targets (e.g., antibodies that bind to other cytokines or growth factors, their receptors, or other cell surface molecules); and anti-inflammatory cytokines or agonists thereof. Non-limiting examples of the agents that can be used in combination with the antibodies described herein, include, but are not limited to, antagonists of one or more interleukins (ILs) or their receptors, e.g., antagonists of IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-13, IL-15, IL-16, IL17A-F, IL-18, IL-20, IL-21, IL-22, IL-25 and IL-31; antagonists of cytokines or growth factors or their receptors, such as, LT, EMAP-II, GM-CSF, FGF and PDGF. Antibodies of the invention can also be combined with inhibitors of e.g., antibodies to, cell surface molecules such as CD2, CD3, CD4, CD8, CD20 (e.g., the CD20 inhibitor rituximab (RITUXAN®), CD25, CD28, CD30, CD40, CD45, CD69, CD80 (B7.1), CD86 (B7.2), CD90, or their ligands, including CD154 (gp39 or CD40L), or LFA-1/ICAM-1 and VLA-4/VCAM-1 (Yusuf-Makagiansar et al.,  Med. Res. Rev.,  22:146-167 (2002)). Preferred antagonists that can be used in combination with one or more antibodies, e.g., bispecific antibodies, described herein include antagonists of IL-1, IL-6, IL-12, TNF-α, IL-15, IL-18, IL-20, IL-22 and IL-31. 
     Examples of those agents include IL-12 antagonists, such as chimeric, humanized, human or in vitro-generated antibodies (or antigen binding fragments thereof) that bind to IL-12 (preferably human IL-12), e.g., the antibody disclosed in WO 00/56772; IL-12 receptor inhibitors, e.g., antibodies to human IL-12 receptor; and soluble fragments of the IL-12 receptor, e.g., human IL-12 receptor. Examples of IL-15 antagonists include antibodies (or antigen binding fragments thereof) against IL-15 or its receptor, e.g., chimeric, humanized, human or in vitro-generated antibodies to human IL-15 or its receptor, soluble fragments of the IL-15 receptor, and IL-15-binding proteins. Examples of IL-17 antagonists include brodalumab, secukinumab, and ixekizumab. Examples of IL-18 antagonists include antibodies, e.g., chimeric, humanized, human or in vitro-generated antibodies (or antigen binding fragments thereof), to human IL-18, soluble fragments of the IL-18 receptor, and IL-18 binding proteins (IL-18BP). Examples of IL-1 antagonists include Interleukin-1-converting enzyme (ICE) inhibitors, such as Vx740, IL-1 antagonists, e.g., IL-1 RA (anakinra, Kineret®), sIL1RII, and anti-IL-1 receptor antibodies (or antigen binding fragments thereof). 
     In other embodiments, one or more antibodies, e.g., bispecific antibodies, described herein may be administered in combination with one or more of the following: IL-13 antagonists, e.g., soluble IL-13 receptors (sIL-13) and/or antibodies against IL-13; IL-2 antagonists, e.g., DAB 486-IL-2 and/or DAB 389-IL-2 (IL-2 fusion proteins, Seragen), and/or antibodies to IL-2R, e.g., anti-Tac (humanized anti-IL-2R, Protein Design Labs). Yet another combination includes one or more antibodies, e.g., bispecific antibodies, of the invention, antagonistic small molecules, and/or inhibitory antibodies in combination with nondepleting anti-CD4 inhibitors (DEC-CE9.1/SB 210396; non-depleting primatized anti-CD4 antibody; IDEC/SmithKline). Yet other preferred combinations include antagonists of the costimulatory pathway CD80 (B7.1) or CD86 (B7.2), including antibodies, soluble receptors or antagonistic ligands; as well as p-selectin glycoprotein ligand (PSGL), anti-inflammatory cytokines, e.g., IL-4 (DNAX/Schering); IL-10 (SCH 52000; recombinant IL-10 DNAX/Schering); IL-13 and TGF-beta, and agonists thereof (e.g., agonist antibodies). 
     In other embodiments, one or more antibodies, e.g., bispecific antibodies, of the invention can be co-formulated with, and/or co-administered with, one or more anti-inflammatory drugs, immunosuppressants, or metabolic or enzymatic inhibitors. Non-limiting examples of the drugs or inhibitors that can be used in combination with the antibodies described herein, include, but are not limited to, one or more of: nonsteroidal anti-inflammatory drug(s) (NSAIDs), e.g., ibuprofen, tenidap, naproxen, meloxicam, piroxicam, diclofenac, and indomethacin; sulfasalazine; corticosteroids such. as prednisolone; cytokine suppressive anti-inflammatory drug(s) (CSAIDs); inhibitors of nucleotide biosynthesis, e.g., inhibitors of purine biosynthesis, folate antagonists (e.g., methotrexate (N-[4-[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-glutamic acid); and inhibitors of pyrimidine biosynthesis, e.g., dihydroorotate dehydrogenase (DHODH) inhibitors. Preferred therapeutic agents for use in combination with one or more antibodies, e.g., bispecific antibodies, of the invention include NSAIDs, CSAIDs, (DHODH) inhibitors (e.g., leflunomide), and folate antagonists (e.g., methotrexate). 
     Examples of additional inhibitors include one or more of: corticosteroids (oral, inhaled and local injection); immunosuppresants, e.g., cyclosporin, tacrolimus (FK-506); and mTOR inhibitors, e.g., sirolimus (rapamycin—RAPAMUNE® or rapamycin derivatives, e.g., soluble rapamycin derivatives (e.g., ester rapamycin derivatives, e.g., CCI-779); agents which interfere with signaling by proinflammatory cytokines such as IL-1 (e.g., IRAK, NIK, IKK, p38 or MAP kinase inhibitors); COX2 inhibitors, e.g., celecoxib, rofecoxib, and variants thereof; phosphodiesterase inhibitors, e.g., R973401 (phosphodiesterase Type IV inhibitor); phospholipase inhibitors, e.g., inhibitors of cytosolic phospholipase 2 (cPLA2) (e.g., trifluoromethyl ketone analogs); inhibitors of vascular endothelial cell growth factor or growth factor receptor, e.g., VEGF inhibitor and/or VEGF-R inhibitor; and inhibitors of angiogenesis. Preferred therapeutic agents for use in combination with the antibodies of the invention are immunosuppresants, e.g., cyclosporin, tacrolimus (FK-506); mTOR inhibitors, e.g., sirolimus (rapamycin) or rapamycin derivatives, e.g., soluble rapamycin derivatives (e.g., ester rapamycin derivatives, e.g., CCI-779); COX2 inhibitors, e.g., celecoxib and variants thereof; and phospholipase inhibitors, e.g., inhibitors of cytosolic phospholipase 2 (cPLA2), e.g., trifluoromethyl ketone analogs. 
     Additional examples of therapeutic agents that can be combined with an antibody, e.g., bispecific antigen binding protein, of the invention include one or more of: 6-mercaptopurines (6-MP); azathioprine sulphasalazine; mesalazine; olsalazine; chloroquine/hydroxychloroquine (PLAQUENIL®); pencillamine; aurothiornalate (intramuscular and oral); azathioprine; coichicine; beta-2 adrenoreceptor agonists (salbutamol, terbutaline, salmeteral); xanthines (theophylline, aminophylline); cromoglycate; nedocromil; ketotifen; ipratropium and oxitropium; mycophenolate mofetil; adenosine agonists; antithrombotic agents; complement inhibitors; and adrenergic agents. 
     Anti-TL1A antibodies of the invention may be combined with TNF-α antagonists for treatment of the same conditions as noted herein for anti-TL1A/anti-TNF-α bispecific antigen binding proteins. Such TNF-α antagonists include, for example, etanercept, adalimumab, infliximab, golimumab, and certolizumab pegol. 
     Non-limiting examples of agents for treating or preventing arthritic disorders (e.g., rheumatoid arthritis, inflammatory arthritis, rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis and psoriatic arthritis), with which an antibody, e.g., bispecific antigen binding protein, of the invention may be combined include one or more of the following: IL-12 antagonists as described herein; NSAIDs; CSAIDs; nondepleting anti-CD4 antibodies as described herein; IL-2 antagonists as described herein; anti-inflammatory cytokines, e.g., IL-4, IL-10, IL-13 and TGF-α, or agonists thereof; IL-1 or IL-1 receptor antagonists as described herein; phosphodiesterase inhibitors as described herein; Cox-2 inhibitors as described herein; iloprost: methotrexate; thalidomide and thalidomide-related drugs (e.g., Celgen); leflunomide; inhibitor of plasminogen activation, e.g., tranexamic acid; cytokine inhibitor, e.g., T-614; prostaglandin E1; azathioprine; an inhibitor of interleukin-1 converting enzyme (ICE); zap-70 and/or 1 ck inhibitor (inhibitor of the tyrosine kinase zap-70 or 1 ck); an inhibitor of vascular endothelial cell growth factor or vascular endothelial cell growth factor receptor as described herein; an inhibitor of angiogenesis as described herein; corticosteroid anti-inflammatory drugs (e.g., SB203580); TNF-α-convertase inhibitors; IL-1; IL-13; IL-17 inhibitors; gold; penicillamine; chloroquine; hydroxychloroquine; chlorambucil; cyclophosphamide; cyclosporine; total lymphoid irradiation; antithymocyte globulin; CD5-toxins; orally administered peptides and collagen; lobenzarit disodium; cytokine regulating agents (CRAs) HP228 and HP466 (Houghten Pharmaceuticals, Inc.); ICAM-1 antisense phosphorothioate oligodeoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TP 10; T Cell Sciences, Inc.); prednisone; orgotein; glycosaminoglycan polysulphate; minocycline (MINOCIN®); anti-IL2R antibodies; marine and botanical lipids (fish and plant seed fatty acids); auranofin; phenylbutazone; meclofenamic acid; flufenamic acid; intravenous immune globulin; zileuton; mycophenolic acid (RS-61443); tacrolimus (FK-506); sirolimus (rapamycin); amiprilose (therafectin); cladribine (2-chlorodeoxyadenosine); and azaribine. Preferred combinations include one or more antibodies, e.g., bispecific antibodies, of the invention in combination with methotrexate or leflunomide, and in moderate or severe rheumatoid arthritis cases, cyclosporine. 
     Preferred examples of inhibitors to use in combination with one or more antigen binding proteins, e.g., bispecific antigen binding proteins, of the invention to treat arthritic disorders include antagonists of IL-12, IL-15, IL-18, IL-22; T cell and B cell-depleting agents (e.g., anti-CD4 or anti-CD22 antibodies); small molecule inhibitors, e.g., methotrexate and leflunomide; sirolimus (rapamycin) and analogs thereof, e.g., CCI-779; cox-2 and cPLA2 inhibitors; NSAIDs; p38 inhibitors, TPL-2, Mk-2 and NFKB inhibitors; RAGE or soluble RAGE; P-selectin or PSGL-1 inhibitors (e.g., small molecule inhibitors, antibodies to PSGL-1, antibodies to P-selectin); estrogen receptor beta (ERB) agonists or ERB-NFKB antagonists. Most preferred additional therapeutic agents that can be co-administered and/or co-formulated with one or more antibodies, e.g., bispecific antibodies, of the invention include one or more of: methotrexate, leflunomide, or a sirolimus (rapamycin) or an analog thereof, e.g., CCI-779. 
     Non-limiting examples of agents for treating or preventing an inflammatory disease or disorder, (e.g., IBD, CD, UC, IBS) with which an antibody, e.g., bispecific antigen binding protein, of the invention can be combined include the following: budenoside; epidermal growth factor; corticosteroids; cyclosporine; sulfasalazine; aminosalicylates; 6-mercaptopurine; azathioprine; metronidazole; lipoxygenase inhibitors; mesalamine; olsalazine; balsalazide; antioxidants; thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1 monoclonal antibodies; anti-IL-6 monoclonal antibodies (e.g., anti-IL-6 receptor antibodies and anti-IL-6 antibodies); growth factors; elastase inhibitors; pyridinyl-imidazole compounds; IL-4, IL-10, IL-13 and/or TGF.beta. cytokines or agonists thereof (e.g., agonist antibodies); IL-11; glucuronide- or dextran-conjugated prodrugs of prednisolone, dexamethasone or budesonide; ICAM-1 antisense phosphorothioate oligodeoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TP10; T Cell Sciences, Inc.); slow-release mesalazine; methotrexate; antagonists of platelet activating factor (PAF); ciprofloxacin; and lignocaine. 
     Non-limiting examples of agents for treating or preventing multiple sclerosis with one or more antibodies, e.g., bispecific antibodies, of the invention can be combined include the following: interferons, e.g., interferon-α (e.g., AVONEX®, Biogen) and interferon-1b (BETASERON®, Chiron/Berlex); Copolymer 1 (Cop-1; COPAXONE®, Teva Pharmaceutical Industries, Inc.); dimethyl fumarate (e.g., BG-12; Biogen); hyperbaric oxygen; intravenous immunoglobulin; cladribine; corticosteroids; prednisolone; methylprednisolone; azathioprine; cyclophosphamide; cyclosporine; cyclosporine A, methotrexate; 4-aminopyridine; and tizanidine. Additional antagonists that can be used in combination with antibodies of the invention include antibodies to or antagonists of other human cytokines or growth factors, for example, LT, IL-1, IL-2, IL-6, EL-7, IL-8, IL-12 IL-15, IL-16, IL-18, EMAP-11, GM-CSF, FGF, and PDGF. Antibodies as described herein can be combined with antibodies to cell surface molecules such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or their ligands. One or more antibodies, e.g., bispecific antibodies, of the invention may also be combined with agents, such as methotrexate, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs, for example, ibuprofen, corticosteroids such as prednisolone, phosphodiesterase inhibitors, adenosine agonists, antithrombotic agents, complement inhibitors, adrenergic agents, agents which interfere with signaling by proinflammatory cytokines as described herein, IL-1b converting enzyme inhibitors (e.g., Vx740), anti-P7s, PSGL, TACE inhibitors, T-cell signaling inhibitors such as kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, azathioprine, 6-mercaptopurines, angiotensin converting enzyme inhibitors, soluble cytokine receptors and derivatives thereof, as described herein, and anti-inflammatory cytokines (e.g., IL-4, IL-10, IL-13 and TGF). 
     Preferred examples of therapeutic agents for multiple sclerosis with which the antibodies of the invention can be combined include dimethyl fumarate (e.g., BG-12; Biogen), interferon-beta, for example, IFN-β-1a and IFN-β-1b; COPAXONE®, corticosteroids, IL-1 inhibitors, antibodies to CD40 ligand and CD80, IL-12 antagonists. 
     Non-limiting examples of agents for treating or preventing psoriasis with which an antibody, e.g., bispecific antigen binding protein, of the invention can be combined include the following: corticosteroids; vitamin. D3 and analogs thereof; retinoiods (e.g., soriatane); methotrexate; cyclosporine, 6-thioguanine; Accutane; hydrea; hydroxyurea; sulfasalazine; mycophenolate mofetil; azathioprine; tacrolimus; fumaric acid esters; biologics such as AMEVIVE®, Raptiva ustekinumab, and XP-828L; phototherapy; and photochemotherapy (e.g., psoralen and ultraviolet phototherapy combined). 
     Non-limiting examples of agents for treating or preventing inflammatory airway/respiratory disease (e.g., chronic obstructive pulmonary disorder, asthma) with which an antibody, e.g., bispecific antigen binding protein, of the invention can be combined include the following: beta2-adrenoceptor agonists (e.g., salbutamol (albuterol), levalbuterol, terbutaline, bitolterol); long-acting beta2-adrenoceptor agonists (e.g., salmeterol, formoterol, bambuterol); adrenergic agonists (e.g., inhaled epinephrine and ephedrine tablets); anticholinergic medications (e.g., ipratropium bromide); combinations of inhaled steroids and long-acting bronchodilators (e.g., fluticasone/salmeterol (ADVAIR® in the United States, and Seretide in the United Kingdom)) or. budesonide/formoterol (SYMBICORT®)); inhaled glucocorticoids (e.g., ciclesonide, beclomethasone, budesonide, flunisolide, fluticasone, mometasone, triamcinolone); leukotriene modifiers (e.g., montelukast, zafirlukast, pranlukast, and zileuton); mast cell stabilizers (e.g., cromoglicate (cromolyn), and nedocromil); antimuscarinics/anticholinergics (e.g., ipratropium, oxitropium, tiotropium); methylxanthines (e.g., theophylline, aminophylline); antihistamines; IgE blockers (e.g., omalizumab); M 3  muscarinic antagonists (anticholinergics) (e.g., ipratropium, tiotropium); cromones (e.g., chromoglicate, nedocromil); zanthines (e.g., theophylline); IL-17 inhibitors (e.g., brodalumab, secukinumab, ixekizumab), IL-4 inhibitors; and IL-13 inhibitors. 
     In one embodiment, an antibody, e.g., bispecific antigen binding protein, of the invention can be used in combination with one or more antibodies directed at other targets involved in regulating immune responses, e.g., transplant rejection. 
     Non-limiting examples of agents for treating or preventing immune responses with which an antibody, e.g., bispecific antigen binding protein, of the invention can be combined include the following: antibodies against other cell surface molecules, including but not limited to CD25 (interleukin-2 receptor-α), CD11a (LFA-1), CD54 (ICAM-1), CD4, CD45, CD28/CTLA4 (CD80 (B7.1), e.g., CTLA4 Ig (abatacept , ORENCIA®), ICOSL, ICOS and/or CD86 (B7.2). In yet another embodiment, an antibody of the invention is used in combination with one or more general immunosuppressive agents, such as cyclosporin A or FK506. 
     In other embodiments, antibodies are used as vaccine adjuvants against autoimmune disorders, inflammatory diseases, etc. The combination of adjuvants for treatment of these types of disorders are suitable for use in combination with a wide variety of antigens from targeted self-antigens, i.e., autoantigens, involved in autoimmunity, e.g., myelin basic protein; inflammatory self-antigens (e.g., amyloid peptide protein) or transplant antigens (e.g., alloantigens). The antigen may comprise peptides or polypeptides derived from proteins, as well as fragments of any of the following: saccharides, proteins, polynucleotides or oligonucleotides, autoantigens, amyloid peptide protein, transplant antigens, allergens, or other macromolecular components. In some instances, more than one antigen is included in the antigenic composition. 
     For example, desirable vaccines for moderating responses to allergens in a vertebrate host, which contain the adjuvant combinations of this invention, include those containing an allergen or fragment thereof. Examples of such allergens are described in U.S. Pat. No. 5,830,877 and PCT Publication No. WO 99/51259, which are hereby incorporated by reference in their entireties, and include pollen, insect venoms, animal dander, fungal spores and drugs (such as penicillin). The vaccines interfere with the production of IgE antibodies, a known cause of allergic reactions. In another example, desirable vaccines for preventing or treating disease characterized by amyloid deposition in a vertebrate host, which contain the adjuvant combinations of this invention, include those containing portions of amyloid peptide protein (APP). This disease is referred to variously as Alzheimer&#39;s disease, amyloidosis or amyloidogenic disease. Thus, the vaccines of this invention include the adjuvant combinations of this invention plus A.beta. peptide, as well as fragments of Aβ peptide and antibodies to Aβ peptide or fragments thereof 
     In another embodiment, pharmaceutical compositions may be supplied as a kit comprising a container that comprises an antibody, bispecific antigen binding protein or antigen-binding fragment of the invention. Antibodies, e.g., bispecific antibodies, of the invention can be provided in the form of an injectable solution for single or multiple doses, or as a sterile powder that will be reconstituted before injection. Alternatively, such a kit can include a dry-powder disperser, liquid aerosol generator, or nebulizer for administration of the antigen binding protein (e.g., anti-TL1A/anti-TNF-α bispecific antigen binding protein). Such a kit may further comprise written information on indications and usage of the pharmaceutical composition. Moreover, such information may include a statement that the antibody composition is contraindicated in patients with known hypersensitivity to TL1A and TNF-α. 
     In a further embodiment, the invention provides an article of manufacture, comprising: (a) a composition of matter comprising an antibody, bispecific antigen binding protein or antigen-binding fragment as described herein; (b) a container containing said composition; and (c) a label affixed to said container, or a package insert included in said container referring to the use of said antibody in the treatment of an immune related disease. 
     In another aspect, the composition comprises a further active ingredient, which may, for example, be a further antibody or an anti-inflammatory, cytotoxic or chemotherapeutic agent. Preferably, the composition is sterile. 
     The antibodies, e.g., bispecific antibodies, as described herein are also useful to prepare medicines and medicaments for the treatment of immune-related and inflammatory diseases, including for example, IBS, IBD, CD, UC. In a specific aspect, such medicines and medicaments comprise a therapeutically effective amount of a bispecific antigen binding protein, antibody or antigen-binding fragment of the invention with a pharmaceutically acceptable carrier. In an embodiment, the admixture is sterile. 
     The bispecific antigen binding proteins of the invention are useful for detecting TL1A and/or TNF-α in biological samples and identification of cells or tissues that express the TL1A receptor DR3 and/or TNF-α receptor. For instance, the bispecific antigen binding proteins can be used in diagnostic assays, e.g., binding assays to detect and/or quantify TL1A and/or TNF-α binding and/or expression in a tissue or cell. In addition, the bispecific antigen binding proteins described herein can be used to inhibit TL1A from forming a complex with its receptor DR3, thereby modulating the biological activity of TL1A or DR3 in a cell or tissue. Likewise, the bispecific antigen binding proteins described herein can be used to inhibit TNF-α from forming a complex with its receptor, thereby modulating the biological activity of TNF-α or its receptor in a cell or tissue. Exemplary activity that can be modulated includes, but is not limited to, inhibiting and/or reducing inflammation. 
     The bispecific antigen binding proteins described herein can be used for diagnostic purposes to detect, diagnose, or monitor diseases and/or conditions associated with TL1A and/or TNF-α. Also provided are methods for the detection of the presence of TL1A and/or TNF-α in a sample using classical immunohistological methods known to those of skill in the art. See, for example, Tijssen (1993),  Practice and Theory of Enzyme Immunoassays,  Vol 15 (Eds R. H. Burdon and P. H. van Knippenberg, Elsevier, Amsterdam); Zola (1987),  Monoclonal Antibodies: A Manual of Techniques,  pp. 147-158 (CRC Press, Inc.); Jalkanen et al. (1985),  J. Cell. Biol.  101:976-985; Jalkanen et al. (1987),  J. Cell Biol.  105:3087-3096. The detection of either TL1A and/or TNF-α can be performed in vivo or in vitro. 
     Diagnostic applications provided herein include use of the antigen binding proteins to detect expression of TL1A and/or TNF-α and binding of these ligands to their receptors. Examples of methods useful in the detection of the presence of the ligand include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). 
     For diagnostic applications, the antigen binding protein typically will be labeled with a detectable labeling group. Suitable labeling groups include, but are not limited to, the following: radioisotopes or radionuclides (e.g.,  3 H,  14 C,  15 N,  35 S,  90 Y,  99 Tc,  111 In,  125 I,  131 I), fluorescent groups (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic groups (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent groups, biotinyl groups, or predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags). In some embodiments, the labeling group is coupled to the antigen binding protein via spacer arms of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and may be used. 
     In another embodiment, the bispecific antigen binding protein described herein can be used to identify a cell or cells that express TL1A and/or TNF-α. In a specific embodiment, the antigen binding protein is labeled with a labeling group and the binding of the labeled antigen binding protein to TL1A and/or TNF-α is detected. In a further specific embodiment, the binding of the antigen binding protein to TL1A and/or TNF-α is detected in vivo. In a further specific embodiment, the bispecific antigen binding protein is isolated and measured using techniques known in the art. See, for example, Harlow and Lane, 1988, Antibodies: A Laboratory Manual, New York: Cold Spring Harbor (ed. 1991 and periodic supplements); John E. Coligan, ed., 1993, Current Protocols In Immunology New York: John Wiley &amp; Sons. 
     Another aspect of the invention provides for detecting the presence of a test molecule that competes for binding to TL1A and/or TNF-α with the antigen binding proteins described herein. An example of one such assay would involve detecting the amount of free antigen binding protein in a solution containing an amount of TL1A and/or TNF-α in the presence or absence of the test molecule. An increase in the amount of free antigen binding protein (i.e., the antigen binding protein not bound to TL1A and/or TNF-α) would indicate that the test molecule is capable of competing for TL1A and/or TNF-α binding with the bispecific antigen binding protein. In one embodiment, the antigen binding protein is labeled with a labeling group. Alternatively, the test molecule is labeled and the amount of free test molecule is monitored in the presence and absence of the antigen binding protein. 
     WORKING EXAMPLES 
     The invention is further illuminated by the following working examples, which exemplify but do not limit the scope of the invention 
     Example 1 
     Preparation of XenoMouse® Anti-TL1A Monoclonal Antibodies 
     Mouse Strains 
     Fully human antibodies to human TL1A were generated by immunizing XENOMOUSE® transgenic mice. U.S. Pat. Nos. 6,114,598; 6,162,963;6,833,268; 7,049,426; 7,064,244, which are incorporated herein by reference in their entirety; Green et al. (1994),  Nature Genetics  7:13-21; Mendez et al. (1997),  Nature Genetics  15:146-156; Green and Jakobovitis (1998),  J. Ex. Med,  188:483-495; Kellerman and Green (2002),  Current Opinion in Biotechnology  13, 593-597; each of which is incorporated by reference in its entirety. Animals from the XMG1-KL, XMG2-K, XMG2-K/Balbc, XMG2-KL, XMG4-K and XMG4-KL XENOMOUSE® strains were used for all immunizations. 
     Generation of TL1A Immunogen 
     TL1A polypeptides containing the N-terminal His tag (H6) of which the first 22 amino acids are the VK1 signal peptide) were generated by transiently transfecting 293HEK cells with the corresponding cDNAs. The commonly used polyHis tag was employed to facilitate detection and subsequent purification. 
     293-6E cells at 9.48×10 5  cells/ml were transfected with 0.5 mg/L DNA (0.1 mg/L His-TL1A in pTTS vector with 0.4 mg/L empty pTTS vector) (Durocher et al. (2002) NRCC,  Nucleic Acids. Res.  30, e9) with 3 ml PEI/mg DNA in FreeStyle 293 media (Invitrogen). Tryptone N1 was added to cultures 1 hour after transfection. Cells were grown in suspension in FreeStyle 293 expression medium supplemented with 0.1% Pluronic F68 and 50 μg/ml Geneticin for 7 days and harvested for purification. 
     Example 2 
     Generation of TL1A 
     TL1A polypeptides containing the N-terminal His tag (H6) of which the first 22 amino acids are the VK1 signal peptide) were generated by transiently transfecting 293HEK cells with the corresponding cDNAs. The commonly used polyHis tag was employed to facilitate detection and subsequent purification. 
     293-6E cells at 9.48×10 5  cells/ml were transfected with 0.5 mg/L DNA (0.1 mg/L His-TL1A in pTT5 vector with 0.4 mg/L empty pTT5 vector) (Durocher et al. NRCC, Nucleic Acids. Res. (2002) 30, e9) with 3m1 PEI/mg DNA in FreeStyle 293 media (Invitrogen). Tryptone N1 was added to cultures 1 hour after transfection. Cells were grown in suspension in FreeStyle 293 expression medium supplemented with 0.1% Pluronic F68 and 50 μg/ml Geneticin for 7 days and harvested for purification. 
     Immunizations 
     Immunizations are conducted using one or more suitable forms of TL1A antigen, including recombinant human TL1A expressed on cells and recombinant human TL1A soluble protein or combinations thereof. 
     A suitable amount of immunogen (i.e., 10 μg of protein delivered by injection to the abdomen) is used for initial immunization in XenoMouse®. Following the initial immunization, subsequent boost immunizations of immunogen (i.e., 2×10 e6 cells or 5 μg of protein) are administered on a schedule and for the duration necessary to induce a suitable titer of anti-TL1A antibody in the mice. Titers are determined by any suitable method, for example, enzyme immunoassay or fluorescence activated cell sorting (FACS). 
     Multiple immunogens and routes of immunization were used to generate anti-human TL1A immune responses. For genetic immunizations, mice were immunized 12-16 times over 6-8 weeks using the Helios Gene Gun system according to the manufacturer&#39;s instructions (BioRad, Hercules, Calif.). Briefly, expression vectors encoding wild type human or cynomolgus TL1A were coated onto gold beads (BioRad, Hercules, Calif.) and delivered to the epidermis of a shaved mouse abdomen. For cell-based immunizations, mice were immunized with a suspension-adapted CHO-K1 cell line (Invitrogen, Carlsbad, Calif.), stably transfected with an expression vector encoding human TL1A. Animals were immunized with cells mixed with Alum prepared from aluminum potassium sulfate (EMD Chemicals Inc., Gibbstown, N.J.) and CpG-ODN (Eurofins MWG Operon LLC, Huntsville, Ala.) 10-12 times over 6-8 weeks using a protocol that alternated between subcutaneous and intraperitoneal injections. The initial boost was comprised of 4×10 6  cells while subsequent boosts contained 2×10 6  cells. For soluble, recombinant protein immunizations, mice were immunized with a 6× His-tagged, trimeric form of the human and cynomolgus TL1A extracellular domain (amino acids 72-251 of the TL1A sequences). Animals were immunized with recombinant protein mixed with Alum and CpG-ODN, 8-12 times over 4-8 weeks using sub-cutaneous injections. The initial boost was comprised of 10 μg while subsequent boosts contained 5 μg. Human TL1A-specific serum titers were monitored by live-cell FACS analysis on an Accuri or FacsCalibur (BD Biosciences) flow cytometer. Animals with the highest antigen-specific serum titers directed against human and cynomolgus TL1A were sacrificed and used for hybridoma generation (Kohler and Milstein, 1975). 
     Example 3 
     Preparation of Monoclonal Antibodies 
     Hybridoma Generation 
     Animals exhibiting suitable titers are identified, and lymphocytes are obtained from draining lymph nodes and, if necessary, pooled for each cohort. Pooled lymphocytes (from each immunization cohort) were dissociated from lymphoid tissue by grinding in a suitable medium (for example, Dulbecco&#39;s Modified Eagle Medium (DMEM); Invitrogen, Carlsbad, Calif.). B cells may be selected and/or expanded using a suitable method, and fused with suitable fusion partner; for example, non-secretory myeloma P3X63Ag8.653 cells (American Type Culture Collection CRL 1580; Kearney et al  J. Immunol.  123, 1979, 1548-1550), using techniques that are known in the art. B cells were selected and/or expanded using standard methods, and fused with a suitable fusion partner using techniques that were known in the art. 
     In one suitable fusion method, lymphocytes are mixed with fusion partner cells at a ratio of 1:4. The cell mixture is gently pelleted by centrifugation at 400×g for 4 minutes, the supernatant decanted, and the cell mixture gently mixed (for example, by using a 1 ml pipette). Fusion is induced with PEG/DMSO (polyethylene glycol/dimethyl sulfoxide; obtainable from Sigma-Aldrich, St. Louis Mo.; 1 ml per million lymphocytes). PEG/DMSO is slowly added with gentle agitation over one minute followed, by one minute of mixing. IDMEM (DMEM without glutamine; 2 ml per million B cells), is then added over 2 minutes with gentle agitation, followed by additional IDMEM (8 ml per million B-cells) which is added over 3 minutes. 
     The fused cells are gently pelleted (400×g, 6 minutes) and resuspended in 20 ml Selection media (for example, DMEM containing Azaserine and Hypoxanthine [HA] and other supplemental materials as necessary) per million B-cells. Cells are incubated for 20-30 minutes at 37 C and then resuspended in 200 ml Selection media and cultured for three to four days in T175 flasks prior to 96-well plating. 
     Cells are distributed into 96-well plates using standard techniques to maximize clonality of the resulting colonies. After several days of culture, supernatants are collected and subjected to screening assays as detailed in the examples below, including confirmation of binding to human TL1A, evaluation of cross-reactivity with other species&#39; TL1A (for example, cynomologous monkey TL1A), and ability to inhibit the activity of TL1A. Positive cells are further selected and subjected to standard cloning and subcloning techniques. Clonal lines may be expanded in vitro, and the secreted human antibodies obtained for analysis. 
     In this manner, mice were immunized with recombinant human TL1A soluble protein for a total of 15 immunizations over a period of approximately 2 months; several hybridoma cell lines secreting TL1A-specific antibodies were obtained, and the antibodies were further characterized. The sequences thereof are presented in the Sequence Listing and in Tables A, C and D and results of various tests using these antibodies are shown herein. 
     Tables A to E herein show the sequences of anti-TL1A antibodies prepared in accordance with this working example. 
     Example 4 
     Antigen Enrichment of Hybridoma Pools 
     Fused hybridoma pools from select immune tissue harvest were used as a source of material for FACS-based enrichments. To enrich for hybridomas expressing antibodies specific to native (full length, on-cell) human TL1A membranes were prepared from 293T cells transiently expressing the TL1A cDNA construct. 24 hours after transfection using 293Fectin™ (ThermoFisher Scientific Inc.) cells were biotinylated with E-Z link NHS-LC-LC-Biotin according to the manufacturer&#39;s recommendation (ThermoFisher Scientific Inc.). After biotinylation, cells were homogenized with a needle and syringe to form membrane fragments and referred to as “membrane preps”. The biotinylated membrane preps were then used to detect hybridomas expressing surface antibodies specific to the target of interest via standard biotin-streptavidin chemistry. 
     To enrich hybridoma pools for the antigen of interest, they were first incubated with the membrane prep probe. Unbound probe was then washed away and the antigen-specific hybridomas were identified by simultaneous detection of surface IgG (with an Alexa 488 conjugated Gt anti-human Fc secondary antibody; Jackson ImmunoResearch) and the biotinylated membrane prep TL1A probe (Alexa Fluor 647 conjugated streptavidin; Jackson ImmunoResearch). Hybridomas expressing surface IgG and binding antigen were detected by FACS analysis on an Accuri flow cytometer. Dual positive events were sorted as single cells into 384-well plates on a FACS Aria cell sorter (BD Biosciences). After several days of culture, the hybridoma supernatants containing monoclonal antibodies were collected and used in the screening assays described in the examples below. 
     Example 5 
     Initial Selection of TL1A-Specific Binding Antibodies 
     Human TL1A was expressed on host Human Embryonic Kidney 293 cells by transfection using an expression vector expressing huTL1A cDNA, Gibco™ Opti-MEMO media (Gibco, Cat. No. 31985088) and 293Fectin™ reagent (Invitrogen, Cat. No. 12347019) following the protocol set out by the manufacturer. Hybridoma supernatants were screened for the presence of huTL1A-specific monoclonal antibodies using the FMAT 8200 Screening System (Molecular Devices) and the Celllnsight™ High Content Imaging Platform (ThermoFisher Scientific). The number of huTL1A positive wells (i.e., those that have signal over irrelevant hybridoma supernatant) is presented in Table 5.1. For CellInsight screens, 15 μl/well of hybridoma supernatant (and positive and negative controls) were added to black, 384-well, clear bottom plates (Corning Can. No. 3712) followed by the addition of 30 μl/well of a mixture of TL1A/293T cells, nuclear Hoescht stain (Pierce, Cat. No. 62249) and Alexa 488-goat anti-human IgG (H+L) (Jackson, Cat. No. 109-545-088). After 3 hours of incubation at room temperature, plates were washed 2 times on an AquaMax 4000 plate washer (fitted with a 384-well cell wash head) and read on the CellInsight instrument according to the manufacturer&#39;s recommendations. For FMAT-based screens, 20 μl/well of hybridoma supernatant (and positive and negative controls) were added to black, 384-well, clear bottom plates followed by addition of 40 μl/well of a mixture of TL1A/293T cells, 293T parental cells and Cy5-goat anti-human IgG (Fc) (Jackson, Cat. No. 109-075). After 3 hours of incubation at room temperature, plates were read on the FMAT 8200 system according to the manufacturer&#39;s recommendations. 
     
       
         
           
               
             
               
                 TABLE 5.1 
               
             
            
               
                   
               
               
                 TL1A-specific antibodies selected 
               
            
           
           
               
               
               
            
               
                 Harvest # 
                 Platform 
                 Total # Positive 
               
               
                   
               
               
                 1 
                 FMAT 8200 
                 1488 
               
               
                 2 
                 FMAT 8200 
                  900 
               
               
                 5 
                 Cellinsight 
                 1830 
               
               
                 6 
                 FMAT 8200 
                  438 
               
               
                 7&amp;8 
                 Cellinsight 
                 1541 
               
               
                   
               
            
           
         
       
     
     Example 6 
     Identification of TL1A Receptor-Ligand Blocking Antibodies 
     Biotinylated huTL1A was prepared by reacting 100 μg/ml of NHS LC LC biotin (Pierce, Cat. No. 21338) and 100 μg huTL1A (prepared as described in Example 2) in 1 ml of PBS pH 8.5 for 1 hr at room temperature. Un-reacted biotin was removed by ultra-filtration using a 5 kDa Amicon Ultra spin column (Millipore, Cat. No. UFC8 005). Hybridoma supernatants containing huTL1A-binding antibodies were assayed for their ability to block huTL1A binding to human Death Receptor 3 (huDR3) via an ELISA-based receptor-ligand assay. ELISA plates (Corning Cat. No. 3702) were coated with 40 μl/well of human DR3-Fc chimera (1 μg/ml) (R&amp;D Systems, Cat. No. 943-D3) in coating buffer (1× PBS/0.05% azide), then incubated overnight at 4° C. Plates were then washed with water 3 times and blocked with 90 μl diluent (1× PBS/1% milk) for 30 min at room temperature. 15 μl of anti-TL1A hybridoma supernatant was preincubated with 45 μl of biotinylated TL1A (30 ng/ml final) in 96-well storage plates (Sigma, Cat. No. P6866) in assay diluent for 2 hr at room temperature prior to adding to the pre-blocked DR3 ELISA plates. Assay plates were then incubated for 1 hr at room temperature. Sample plates were subsequently washed 3 times followed by the addition of 40 μl/well of streptavidin-HRP (Pierce, Cat. No. 21126) and another 1 hr incubation at room temperature. Plates were washed an additional three times and 40 μl TMB (Neogen, Cat. No. 308177) was added. The TMB reaction was incubated for 30 min at room temperature and then quenched with 40 μl/well of 1 N hydrochloric acid. Finally, plates were read on an ELISA plate reader at a wavelength of 450 nm. Cutoffs were set at &lt;38% of signal of negative control, irrelevant ESNs (exhausted supernatants). The numbers of samples able to block the huTL1A-DR3 interaction (as defined by this cutoff) are indicated in Table 6.1. 
     
       
         
           
               
             
               
                 TABLE 6.1 
               
             
            
               
                   
               
               
                 Selected TL1A/DR3 blockers 
               
            
           
           
               
               
               
            
               
                   
                 Harvest # 
                 Total #TL1A/DR3 blockers 
               
               
                   
                   
               
               
                   
                 1 
                 208 
               
               
                   
                 2 
                  39 
               
               
                   
                   
               
            
           
         
       
     
     Example 7 
     TL1A Functional Blocking Assays 
     In order to screen for hybridomas capable of blocking TL1A functional activity, IFNγ release from primary T cells or a NF-κB reporter assay in TF1 cells were employed. For the IFNγ release assay, purified primary human T cells (Biological Speciality Corp.,Cat. #215-01-10) were stimulated with soluble human TL1A in the presence or absence of hybridoma supernatants specific to TL1A. 2×10 5  primary human T cells were stimulated with 8-16 ng/mL human TL1A (Amgen), 1-2 ng/mL IL-12 (Peprotech) and 0.5-1 ng/mL IL-18 (R&amp;D Systems) in the presence of hybridoma supernatants containing anti-TL1A antibodies in 96-well round bottom plate at 37° C. for 72 hours. Culture supernatants were then tested for IFNγ level by ELISA according to the manufacturer&#39;s instructions (R&amp;D Systems). For the TL1A responsive reporter assay, a TF-1 NF-κB reporter cell line (Amgen) was stimulated with soluble or membrane-bound human TL1A or soluble cynomolgus monkey TL1A. 0.2-3 nM or 2-20 nM of soluble human or cynomolgus monkey TL1A (respectively) was incubated with 10 4 -10 5  TF-1 NF-KB reporter cells in the presence of serially diluted hybridoma supernatants (or controls) in 96 or 384-well plates at 37° C. overnight. For testing samples against membrane-bound TL1A, activity assays were performed by co-culturing the TF-1 NF-KB reporter cell line with human TL1A-expressing AMID cells. 10 5  TF-1 NF-KB reporter cells and 10 3  AM1D cells were co-cultured in the presence of 5 ug/mL of anti-TL1A antibody (or controls) in a 384-well plate at 37° C. overnight. Reporter signal in each well was determine using the Steady-Glo Luciferase Assay System according to the manufacturer&#39;s recommendation (Promega). 
     Example 8 
     Molecular Rescue and Sequencing of TL1A Receptor-Ligand Blocking Antibodies 
     RNA (total or mRNA) was purified from wells containing the TL1A-neutralizing antibody-producing hybridoma cells using a Qiagen RNeasy mini or the Invitrogen mRNA catcher plus kit. Purified RNA was used to amplify the antibody heavy and light chain variable region (V) genes using cDNA synthesis via reverse transcription, followed by a polymerase chain reaction (RT-PCR). The fully human antibody gamma heavy chain was obtained using the Qiagen One Step Reverse Transcriptase PCR kit (Qiagen). This method was used to generate the first strand cDNA from the RNA template and then to amplify the variable region of the gamma heavy chain using multiplex PCR (see Table 8.1 for the complete primer list, SEQ ID NOS: 1191 to 1252, respectively). The 5′ gamma chain-specific primer annealed to the signal sequence of the antibody heavy chain, while the 3′ primer annealed to a region of the gamma constant domain. The fully human kappa light chain was obtained using the Qiagen One Step Reverse Transcriptase PCR kit (Qiagen). This method was used to generate the first strand cDNA from the RNA template and then to amplify the variable region of the kappa light chain using multiplex PCR. The 5′ kappa light chain-specific primer annealed to the signal sequence of the antibody light chain while the 3′ primer annealed to a region of the kappa constant domain. The fully human lambda light chain was obtained using the Qiagen One Step Reverse Transcriptase PCR kit (Qiagen). This method was used to generate the first strand cDNA from the RNA template and then to amplify the variable region of the lambda light chain using multiplex PCR. The 5′ lambda light chain-specific primer annealed to the signal sequence of light chain while the 3′ primer annealed to a region of the lambda constant domain. 
     The amplified cDNA was purified enzymatically using exonuclease I and alkaline phosphatase and the purified PCR product was sequenced directly. Amino acid sequences were deduced from the corresponding nucleic acid sequences bioinformatically. Two additional, independent RT-PCR amplification and sequencing cycles were completed for each hybridoma sample in order to confirm that any mutations observed were not a consequence of the PCR. The derived amino acid sequences were then analyzed to determine the germline sequence origin of the antibodies and to identify deviations from the germline sequence. The amino acid sequences corresponding to CDRs of the sequenced antibodies were aligned and these alignments were used to group the clones by similarity. 
     
       
         
           
               
             
               
                 TABLE 8.1 
               
             
            
               
                   
               
               
                 Multiplex primers used to amplify 
               
               
                 antibody V genes. 
               
            
           
           
               
               
               
               
            
               
                   
                   
                   
                 SEQ 
               
               
                   
                 Heavy chain 
                 Sequence 5’ to 3’ 
                 ID NO. 
               
               
                   
               
               
                   
                 5&#39; primer 
                 C ACC ATG GAC TGG 
                 1191 
               
               
                   
                   
                 ACC TGG AGG ATC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC TGG 
                 1192 
               
               
                   
                   
                 ACC TGG AGC ATC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC TGC 
                 1193 
               
               
                   
                   
                 ACC TGG AGG ATC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC TGG 
                 1194 
               
               
                   
                   
                 ACC TGG AGA ATC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC TGG 
                 1195 
               
               
                   
                   
                 ACC TGG AGG G 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC TGG 
                 1196 
               
               
                   
                   
                 ATT TGG AGG ATC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC ACA 
                 1197 
               
               
                   
                   
                 CTT TGC TCC ACG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC ACA 
                 1198 
               
               
                   
                   
                 CTT TGC TAC ACA 
                   
               
               
                   
                   
                 CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAG TTT 
                 1199 
               
               
                   
                   
                 GGG CTG AGC TG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA TTG 
                 1200 
               
               
                   
                   
                 GGG CTG AGC TG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAG TTG 
                 1201 
               
               
                   
                   
                 GGG CTG AGC TG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA CTG 
                 1202 
               
               
                   
                   
                 GGG CTC CGC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA TTT 
                 1203 
               
               
                   
                   
                 GGG CTG AGC TGG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAG TTG 
                 1204 
               
               
                   
                   
                 GGG CTG TGC TG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAG TTT 
                 1205 
               
               
                   
                   
                 GGG CTT AGC TGG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAG TTT 
                 1206 
               
               
                   
                   
                 TGG CTG AGC TGG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG AAA CAC 
                 1207 
               
               
                   
                   
                 CTG TGG TTC TTC 
                   
               
               
                   
                   
                 CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG AAG CAC 
                 1208 
               
               
                   
                   
                 CTG TGG TTC TTC 
                   
               
               
                   
                   
                 C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG AAA CAT 
                 1209 
               
               
                   
                   
                 CTG TGG TTC TTC 
                   
               
               
                   
                   
                 CTT CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GGG TCA 
                 1210 
               
               
                   
                   
                 ACC GCC ATC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG TCT GTC 
                 1211 
               
               
                   
                   
                 TCC TTC CTC ATC TTC 
                   
               
               
                   
               
               
                   
                 3&#39; primer 
                 GCTGAGGGAGTAGAGTC 
                 1212 
               
               
                   
                   
                 CTGAGGACTGT 
               
               
                   
               
               
                   
                   
                   
                 SEQ 
               
               
                   
                 Kappa chain 
                 Sequence 5’ to 3’ 
                 ID NO. 
               
               
                   
               
               
                   
                 5&#39; primer 
                 C ACC ATG GAC ATG 
                 1213 
               
               
                   
                   
                 AGG GTC CCC G 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC ATG 
                 1214 
               
               
                   
                   
                 AGG GTC CCT GC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAC ATG 
                 1215 
               
               
                   
                   
                 AGG GTC CTC GC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG AGG CTC 
                 1216 
               
               
                   
                   
                 CCT GCT CAG C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG AGG CTC 
                 1217 
               
               
                   
                   
                 CTT GCT CAG CTT C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA ACC 
                 1218 
               
               
                   
                   
                 CCA GCG CAG C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA GCC 
                 1219 
               
               
                   
                   
                 CCA GCG CAG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA GCC 
                 1220 
               
               
                   
                   
                 CCA GCT CAG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GAA CCA 
                 1221 
               
               
                   
                   
                 TGG AAG CCC CAG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GTG TTG 
                 1222 
               
               
                   
                   
                 CAG ACC CAG GTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GGG TCC 
                 1223 
               
               
                   
                   
                 CAG GTT CAC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG TTG CCA 
                 1224 
               
               
                   
                   
                 TCA CAA CTC ATT GGG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GTG TCC 
                 1225 
               
               
                   
                   
                 CCG TTG CAA TTC 
                   
               
               
                   
               
               
                   
                 3&#39; primer 
                 ACCCGATTGGAGGGCGT 
                 1226 
               
               
                   
                   
                 TATCCACC 
               
               
                   
               
               
                   
                   
                   
                 SEQ 
               
               
                   
                 Lambda chain 
                 Sequence 5’ to 3’ 
                 ID NO. 
               
               
                   
               
               
                   
                 5&#39; primer 
                 C ACC ATG GCC TGG 
                 1227 
               
               
                   
                   
                 TCC CCT CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1228 
               
               
                   
                   
                 TCT CCT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC AGC 
                 1229 
               
               
                   
                   
                 TTC CCT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC GGC 
                 1230 
               
               
                   
                   
                 TTC CCT CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG ACC TGC 
                 1231 
               
               
                   
                   
                 TCC CCT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1232 
               
               
                   
                   
                 GCT CTG CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1233 
               
               
                   
                   
                 GCT CTG CTG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCA TGG 
                 1234 
               
               
                   
                   
                 ATC CCT CTC TTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1235 
               
               
                   
                   
                 ACC GCT CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1236 
               
               
                   
                   
                 ACC CCT CTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1237 
               
               
                   
                   
                 ATC CCT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1238 
               
               
                   
                   
                 ACC GTT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCA TGG 
                 1239 
               
               
                   
                   
                 GCC ACA CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1240 
               
               
                   
                   
                 ATC CCT CTA C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1241 
               
               
                   
                   
                 GTC TCC TTC TAC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1242 
               
               
                   
                   
                 ACC CAA CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCT TGG 
                 1243 
               
               
                   
                   
                 ACC CCA CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1244 
               
               
                   
                   
                 ACT CCT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1245 
               
               
                   
                   
                 ACT CCT CTT CTT C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1246 
               
               
                   
                   
                 ACT CTT CTC CTTC 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1247 
               
               
                   
                   
                 GCT CCA CTA C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1248 
               
               
                   
                   
                 ACT CCT CTC TTT C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1249 
               
               
                   
                   
                 ATG ATG CTT CTC C 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG GCC TGG 
                 1250 
               
               
                   
                   
                 GCT CCT CTG 
                   
               
               
                   
               
               
                   
                   
                 C ACC ATG CCC TGG 
                 1251 
               
               
                   
                   
                 GCT CTG CTC 
                   
               
               
                   
               
               
                   
                 3&#39; primer 
                 GGA GGG TKT GGT 
                 1252 
               
               
                   
                   
                 GGT CTC CAC TCC 
                   
               
               
                   
                   
                 C 
               
               
                   
               
               
                 (where K = G + T) 
               
            
           
         
       
     
     Example 9 
     Preparation of Hetero Ig Constructs 
     Generation of a bispecific antibody through co-expression of two different antibodies leads to contaminants primarily consisting of mispaired heavy and light chains. The preferred bispecific, heterotetramer molecule with two different heavy chains associated with correctly paired light chains is only a minority of the total amount of combinations that can assemble. The contaminants occur mainly due to two different reasons. The first reason is that the heavy chain that comes together at the Fc region of the antibody can homodimerize, leading to conventional monospecific antibody, or heterodimerize, leading to a potential bispecific antibody. The second reason is that light chain is promiscuous and can pair with either of the heavy chains, leading to mispaired light-heavy chain Fab assembly that may not retain binding to the desired target. For these reasons, the bispecific engineering is a two-step process. The first goal is to prevent the homodimerization of the heavy chains and encourage heterodimerization. This can be achieved through engineering the Fc region of the antibodies, using, for example, the knobs-into-holes or charge pair mutations strategies. The second goal is to engineer the light-heavy chain interface in such a way that the light chain is specifically associated only with its cognate heavy chain. 
     The “Hetero-Ig” platform technology (see, e.g., WO2009089004 and W02014081955, both of which are hereby incorporated by reference in their entireties) takes advantage of the electrostatic steering mechanism to overcome the pairing problems mentioned above. Specifically, charged residues are introduced or exploited to drive heavy chain heterodimerization and correct light-heavy chain association. The charge pair mutations (CPMs) in the CH3 domain of the Fc region drive the heterodimerization of the two different heavy chains through opposite charges that cause electrostatic attraction (see, e.g., WO2009089004 and U.S. Pat. No. 8,592,562); the two identical heavy chain combinations have identical juxtaposed charges and are therefore repelled. 
     The correct heavy chain-light chain pairing is facilitated by CPMs at the HC/LC binding interface or the HC1/HC2 binding interface (see  FIG. 1 ). The correct heavy chain-light chain combinations will have opposite charges and therefore be attracted to each other, whereas the incorrect heavy chain-light chain combinations will have the same charges juxtaposed, resulting in repulsion. In  FIG. 1 , correctly assembled hetero-Ig molecules have two or three HC1/HC2 CPMs and two to four HC/LC CPMs that drive the assembly of the preferred heterotetramer comprising two different heavy chains and two different light chains so that the heterotetramer will be the majority component generated by the expression system. The DNAs encoding anti-TL1A/anti-TNF-α hetero Ig-s contain fragments coding for anti-TL1A (or anti-TNF-a) heavy chain and anti-TNF-α (or anti-TL1A) Fab. The DNAs were cloned into pTT5.1 vector. These expression vectors were then used to transfect and express anti-TL1A/anti-TNF-α bi-specifics in human 293 6E cells. 
     Anti-TNF-α antibodies 3.2, 234 and certolizumab were used with anti-TL1A antibodies 3B3, 2G11, 23B3 VH3, 23B3 VH4, and 3C6 to engineer hetero Ig molecules as described in Table J using high throughput cloning, expression and purification. Each of the bispecific hetero Ig molecules had one of the four formats shown in  FIG. 1  using the IgG1 effector functionless scaffold or an IgG2 scaffold. Preferred IgG molecules incorporate the charge mutations shown in  FIG. 1  (v 2 ), which are shown in Table M. The IgG1 effector functionless scaffold comprises substitutions R292C and V302C and may also comprise substitution N297G (also known as the SEFL2 scaffold). 
     Example 10 
     Method for Expression and Purification of anti-TL1A/anti-TNFα Hetero-Ig Molecules 
     Hetero-Ig expression was performed via transient transfection of 293-6E cells. One day prior to transfection N-1 culture was set up in a 20 L Wave bag, at 36° C.-37° C., 5% CO 2 , 0.2 LPM overlay with a total volume of 9 L of culture at 8.5 E5 vc/mL in Freestyle F-17 media (Thermo Fisher). The transfection complex was then prepared by mixing FreeStyle F-17 media, pre-warmed with 0.5 mg/L transfection DNA, with a 1:1:1:1 plasmid DNA chain ratio. Transfection complex (media+DNA+PEI reagent) volume was 10% of final culture volume. Four hours post-transfection , a feed of yeastolate and glucose was added. Culture was harvested on day six at 2.11 E6 cells/mL and 78.2% viability. Expression titer was measured using the ForteBio Octet Q System at 84.0 mg/L. The conditioned media was harvested by centrifugation and filtered using 0.2 μm cellulose acetate filter using a peristaltic pump. 
     For purification, the hetero-Ig molecules were affinity captured by MabSelect SuRe chromatography (GE Life Sciences, Piscataway, N.J.), using Dulbecco&#39;s PBS without divalent cations (Invitrogen, Carlsbad, Calif.) as the wash buffer and 100 mM acetic acid, pH 3.6 as the elution buffer ( FIG. 3 ). All separations were carried out at ambient temperature. The elution peak was pooled based on the chromatogram, neutralized to pH 7.0 using 2 M tris base, diluted with 5-volumes water, and filtered through a 0.22 μm cellulose acetate filter. To remove half antibody species, the sample was then loaded on to an SP-HP sepharose column (GE Life Sciences, Piscataway, N.J.) and washed with 8 column volumes of SP-Buffer A (20 mM sodium phosphate, pH 7.0) followed by elution using a 20 column volume gradient to 60% SP-Buffer B (20 mM sodium phosphate, 1 M NaCl, pH 7.0) ( FIG. 4 ). A pool was made based on the chromatogram and Caliper LabChip (Perkin Elmer, Waltham, Mass.) analysis of fractions. The pool was conditioned with an equal volume of 2× HIC Buffer (200 mM sodium phosphate, 1.5 M ammonium sulfate, pH 7.0) and filtered through a 0.22 p.m cellulose acetate filter. To remove mispaired species, the sample was loaded on to a Butyl-HP sepharose column (GE Life Sciences, Piscataway, N.J.) and washed with 8 column volumes of Butyl-Buffer A (50 mM sodium phosphate, 0.75 M ammonium sulfate, pH 7.0) followed by elution using a 20 column volume gradient to 100% Butyl-Buffer B (50 mM sodium phosphate, pH 7.0) ( FIG. 5 ). A pool was made based on the chromatogram and Caliper LabChip (Perkin Elmer, Waltham, Mass.) analysis of fractions under non-reducing and reducing conditions. The pool was diafiltered against approximately 30 volumes of 10 mM sodium acetate, 9% sucrose, pH 5.2 using Slide-A-Lyzer dialysis cassettes with a 10 kDa cutoff membrane (Pierce, Rockford, Ill.) and further concentrated using a Vivaspin-20 centrifugal concentrator with a 10 kDa cutoff membrane (Sartorius Stedim Biotech, Goettingen, Germany). The concentrated material was then filtered through a 0.8/0.2 μm cellulose acetate filter and the concentration was determined by the absorbance at 280 nm using an extinction coefficient of approximately 212,000. Sample purity was determined by Caliper LabChip analysis under reducing (with 2% 2-mercaptoethanol) and non-reducing (with 25 mM iodoacetamide) conditions ( FIG. 6 ). Analytical SEC was carried out using a Zenix-C SEC-300 column (Sepax Technologies, Newark, Del.) with an isocratic elution in 50 mM sodium phosphate, 250 mM NaCl, pH 6.9 over 18′ ( FIG. 7 ). 
     In order to assess both hetero-Ig integrity and confirm heavy chain-light chain pairing, LC-MS was performed on non-reduced hetero-Ig, as well as hetero-Ig after limited lysyl endoproteinase C (Wako, Richmond, Va.) digestion to produce the hetero-Ig Fab&#39;s (fragment antigen binding) regions of the hetero-Ig. Analysis of non-reduced hetero-Ig was performed by simple dilution of 30 μg native hetero-Ig sample, 1:1 in 0.1% TFA, and injecting 20 pg. Analysis of Fab&#39;s was achieved by incubation of hetero-Ig in the presence of lysyl endoproteinase C using a 1:400 enzyme/substrate ratio in the presence of 100 mM Tris, adjusted to pH8, for 30 minutes at 37° C. The reaction was then stopped by dilution in an equal volume of 0.1% TFA. Initial cleavage of antibody by Lysyl Endoproteinase C is just above the hinge disulfides after the lysine in the sequence motif SCDK/THTCPPC yielding Fab and Fc fragments. 
     Mass analysis was performed using an Agilent 6230 ESI-TOF Mass Spectrometer and 1260 quaternary HPLC system equipped with a Zorbax 300SB-C8, 2.1×50 mm 3.5 μm column (Agilent, Santa Clara, Calif.). Mobile phase A consisted of 0.1% TFA, and mobile phase B consisted of 90% n-propanol, 0.1% TFA in water. Chromatographic gradient conditions for analysis of non-reduced hetero-Ig were as follows: 20% mobile phase B for 1 minute; 1-9 min, 20-70% B; 9-10 min, 70-100% B; 10-11 min, 100% B. Column temperature was kept at 75° C. and post column equilibration at 20% mobile phase B was performed for 7 minutes prior to injection of the next sample. The ESI-TOF settings were as follows: capillary voltage, 5900 V; gas temperature, 340° C.; dry gas, 13 L/min; nebulizer pressure, 25 psig; fragmentor voltage 460 V and skimmer voltage, 95 V ( FIG. 8 ). 
     Analysis of hetero-Ig digested with lysl endoproteinase C was conducted by injecting 20 μg onto the reverse-phase HPLC column and eluting using the following LC gradient: 2% mobile phase held for 2 minutes; 2-12 min, 2-45% B; 12-16 min, 45-90% B; 16-17 min 90% B. Column temperature was kept at 75° C. and post column equilibration at 20% mobile phase B was performed for 7 minutes prior to injection of the next sample ( FIG. 9 ). 
     Example 11 
     Preparation of IgG-ScFv Constructs 
     Each anti-TL1A/anti-TNF-α IgG-scFv antigen binding protein consists of two antigen binding domains, one directed against TL1A and the other against TNF-α. The DNAs encoding anti-TL1A/anti-TNF-α IgG-scFv contain fragments coding for anti-TL1A (or ant-TNF-α) heavy chain (HC) in which the C-terminus is fused to anti-TNF-α (or anti-TL1A) antibody single chain Fv (scFv) (see  FIG. 2 ) with or without cysteine clamp for the purpose of improving biophysical properties. In order to introduce the cysteine clamp, positions 44 (Kabat numbering) VH and 100 (Kabat numbering) in VL were mutated to cysteine. The DNAs were cloned into pTT5.1 vector. These expression vectors were then used to transfect and express anti-TL1A/anti-TNF-α bi-specifics in human 293-6E cells. Full sequences for anti-TL1A/anti-TNF-α IgG-scFv antigen binding proteins produced are shown in Tables L and M. 
     Example 12 
     Method for Purifying anti-TL1A/Anti-TNFα IgG-scFv Molecules 
     IgG-ScFv expression was performed via transient 293 productions. One day prior to transfection, cultures were set up in eight 5 L Thompson Ultra Yield flasks, with a total volume of 2.250 L of culture at 8.5E5 vc/mL each in Freestyle F-17 media (Thermo Fisher). The cultures were kept at 36° C.-37° C., 5% CO 2  at and shaking at 120 RPM. The transfection complex was prepared by mixing FreeStyle F-17 media with 0.5 mg/L DNA, using a 20% of coding plasmid and 80% empty pTT5 vector and PEI at 10% final culture volume Four hours later a yeast lysate and glucose feed was added to each flask. Six days post transfection the cells were harvested through centrifugation, pooled and filtered. Average titer was measured by Forte Bio Octet at 25 mg/L. 
     The IgG-scFv molecules were affinity captured by MabSelect SuRe chromatography (GE Life Sciences, Piscataway, N.J.), using Dulbecco&#39;s PBS without divalent cations (Invitrogen, Carlsbad, Calif.) as the wash buffer and 100 mM acetic acid, pH 3.6 as the elution buffer ( FIG. 10 ). Affinity separations were carried out at ambient temperature. The elution peak was pooled based on the chromatogram, neutralized to pH 7.0 using 2 M tris base, diluted with one volume 10 mM citrate, 75 mM lysine, 4% trehalose, pH 7.0, and concentrated to approximately 20 mg/mL using a Vivacell-100 centrifugal concentrator with a 30 kDa cutoff membrane (Sartorius Stedim Biotech, Goettingen, Germany). To remove high molecular weight aggregates, the sample was filtered through a 0.8/0.2-μm cellulose acetate filter then loaded on to a Superdex 200 Prep Grade column (GE Life Sciences, Piscataway, N.J.) equilibrated with 10 mM citrate, 75 mM lysine, 4% trehalose, pH 7.0 and eluted with an isocratic gradient with the same buffer ( FIG. 11 ). Gel filtration separations were carried out at approximately 7° C. A pool was made based on the chromatogram and analytical SEC of the fractions using a Zenix-C SEC-300 column (Sepax Technologies, Newark, Del.). The pool was further concentrated using a Vivaspin-20 centrifugal concentrator with a 30 kDa cutoff membrane (Sartorius Stedim Biotech, Goettingen, Germany) and filtered through a 0.8/0.2-μm cellulose acetate filter. The concentration was determined by the absorbance at 280 nm using an extinction coefficient of approximately 341,000. Sample purity was determined by Caliper LabChip analysis under reducing (with 2% 2-mercaptoethanol) and non-reducing (with 25 mM iodoacetamide) conditions ( FIG. 12 ). Analytical SEC was carried out using a Zenix-C SEC-300 column (Sepax Technologies, Newark, Del.) with an isocratic elution in 50 mM sodium phosphate, 250 mM NaCl, pH 6.9 over 18′ ( FIG. 13 ). 
     Due to the large size of the Ig-scFvs, 200 kDa or greater, accurate mass was not achievable by ESI-TOF mass analysis. In order to verify mass and assess the quality, Ig-scFv&#39;s were analyzed after digestion with IdeS protease (Promega, Madison Wis.), which cleaves just below the IgG hinge disulfides, between glycines in the sequence motif, CPPCPAPELLG/GP yielding (Fab) 2  and Fc with C-terminally fused scFv. The Ig-scFv&#39;s were incubated in the presence of IdeS protease using a 1:10 enzyme substrate ratio for 1 hour at 37° C. The sample was then diluted 1:1 in 0.1% TFA, and 20 μg was injected onto LC-MS. 
     Mass analysis was performed using an Agilent 6230 ESI-TOF Mass Spectrometer and 1260 quaternary HPLC system equipped with a Zorbax 300SB-C8, 2.1×50 mm 3.5 μm column (Santa Clara, Calif.). Mobile phase A consisted of 0.1% TFA, and mobile phase B consisted of 90% n-propanol, 0.1% TFA in water. Analysis of Ig-scFv digested with IdeS Protease was conducted by injecting 20 μg and eluting using the following LC gradient: 2% mobile phase held for 2 minutes; 2-12 min, 2-45% B; 12-16 min, 45-90% B; 16-17 min 90% B. Column temperature was kept at 75° C. and post column equilibration at 20% mobile phase B was performed for 7 minutes prior to injection of the next sample ( FIG. 14 ). 
     Example 13 
     TL1A Binding Assay 
     Goat anti-human Fc (Jackson Lab, Cat #109-005-098) was first immobilized on an SCMS sensor chip using amine coupling. Anti-TL1A monoclonal antibodies, Fab fragment (using anti-huFab antibody, GE Healthcare cat #28-9583-25) or anti-TL1A/anti-TNF-α bispecific molecules were injected to the chip at 10 μl/min for 1 minute. Various concentrations of human or cynomolgus TL1A protein from 0.78 to 25 nM in sample buffer (PBS, 0.005% P20, 0.1 mg/ml BSA) were injected at flow rate of 50 μl/min for 3 min association, 10 min dissociation. On rate, off rate and equilibrium dissociation constant were calculated using 1:1 binding model on BIAevaluation software. 
     Tables 13.1-13.3 show anti-TL1A binding data for anti-TL1A antibodies, hetero Ig bispecific antibodies, and IgG-scFv bispecific antibodies. 
     
       
         
           
               
             
               
                 TABLE 13.1 
               
             
            
               
                   
               
               
                 TL1A binding of anti-TL1A antibodies 
               
            
           
           
               
               
               
               
               
            
               
                 Anti-TL1A 
                   
                 SEQ ID  
                 human TL1A 
                 cyno TL1A 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 mAb 
                 Protein ID 
                 NOS. 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
               
               
                   
               
               
                 3C6 
                 PL-39112 
                 50, 52 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
                 5.7E+05 
                 1.4E−03 
                 2.4E−09 
               
               
                 3B3 parental 
                 PL-39114 
                 66, 68 
                 4.0E+05 
                  9.5E−05 
                  2.4E−10 
                 2.5E+05 
                 1.4E−04 
                 5.7E−10 
               
               
                 5G4 
                 PL-39115 
                 455, 457 
                 5.0E+05 
                  1.8E−04 
                  3.6E−10 
                 2.9E+05 
                 1.9E−04 
                 6.6E−10 
               
               
                 17E9 
                 PL-39113 
                 459, 461 
                 2.5E+05 
                  5.3E−05 
                  2.2E−10 
                 7.3E+05 
                 2.0E−03 
                 2.8E−09 
               
               
                 9C8 
                 PL-39116 
                 58, 60 
                 1.6E+05 
                  5.3E−05 
                  3.4E−10 
                 9.4E+04 
                 6.9E−05 
                 7.3E−10 
               
               
                 23B3 
                 PL-36543 
                 62, 64 
                 4.7E+05 
                  4.2E−04 
                  8.9E−10 
                 2.3E+05 
                 3.2E−04 
                 1.4E−09 
               
               
                 2G11 
                 PL-36544 
                 54, 56 
                 1.7E+05 
                  1.8E−04 
                  1.1E−09 
                 6.6E+04 
                 4.9E−05 
                 7.3E−10 
               
               
                 3B3 variant 
                 PL-36552 
                 130, 134 
                 4.3E+05 
                  2.2E−04 
                  5.0E−10 
                 2.8E+05 
                 1.4E−04 
                 5.0E−10 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13.2 
               
             
            
               
                   
               
               
                 TL1A binding of anti-TL1A Fab fragment 
               
            
           
           
               
               
               
               
            
               
                 Anti-TL1A  
                   
                 human TL1A 
                 cyno TL1A 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Fab 
                 Protein ID 
                 ka (1/Ms)  
                 kd (1/s) 
                 KD (M) 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 3C6 
                 PL-39193 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
                 NB up to 100 nM 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 3B3 parental  
                 PL-39195 
                 4.1E+05 
                  1.6E−04 
                  4.0E−10 
                 3.5E+05 
                  1.8E−04 
                  5.1E−10 
               
               
                 5G4 
                 PL-39196 
                 4.7E+05 
                  3.3E−04 
                  7.0E−10 
                 4.4E+05 
                  1.6E−04 
                  3.7E−10 
               
            
           
           
               
               
               
               
               
               
            
               
                 17E9 
                 PL-39194 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
                 NB up to 100 nM 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 9C8 
                 PL-39197 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
               
               
                 2363 
                 PL-38024 
                 7.6E+05 
                  6.4E−04 
                  8.5E−10 
                 5.3E+05 
                  1.6E−04 
                  3.1E−10  
               
               
                 2G11 
                 PL-38025 
                 NA 
                 &lt;5.0E−05 
                 &lt;3.0E−10 
                 NA 
                 &lt;5.0E−05 
                 &lt;1.0E−10 
               
               
                 3B3 variant 
                 PL-38026 
                 3.8E+05 
                  4.1E−04 
                  1.1E−09 
                 4.5E+05 
                  5.6E−05 
                  1.3E−10 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13.3 
               
             
            
               
                   
               
               
                 TL1A binding of hetero Ig bispecific antibodies 
               
            
           
           
               
               
               
               
               
               
            
               
                 iPS 
                 anti-TNF 
                 anti-TL1A 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
               
               
                   
               
               
                 376541 
                 Certolizumab 
                 3B3 variant 
                 7.1E+05 
                 2.1E−04 
                 3.0E−10 
               
               
                   
                 parent 
                 (322520) 
                   
                   
                   
               
               
                 376542 
                 C234 
                 3B3 variant 
                 7.1E+05 
                 2.1E−04 
                 3.0E−10 
               
               
                   
                   
                 (322520) 
                   
                   
                   
               
               
                 376543 
                 Certolizumab 
                 3B3 variant 
                 7.2E+05 
                 2.3E−04 
                 3.2E−10 
               
               
                   
                 variant 
                 (322520) 
                   
                   
                   
               
               
                 349461 
                 Certolizumab 
                 2G11 Variant 
                 5.4E+05 
                 1.3E−04 
                 2.5E−10 
               
               
                   
                 Variant 
                   
                   
                   
                   
               
               
                 349463 
                 Certolizumab 
                 23B3 VH4 
                 6.3E+05 
                 4.0E−04 
                 6.4E−10 
               
               
                   
                 Variant 
                   
                   
                   
                   
               
               
                 371222 
                 3.2/CC 
                 3B3 variant 
                 6.6E+05 
                 1.4E−04 
                 2.1E−10 
               
               
                   
                   
                 (322520) 
               
               
                   
               
            
           
         
       
     
     Example 14 
     TL1A Activity Assay 
     TL1A activity assay was performed using TF-1 NF-κB reporter cell line. In brief, 30 ng/ml (EC90) of human or cynomolgus monkey TL1A was incubated with 10 4  TF-1 NF-KB reporter cells in the presence of serially diluted anti-TL1A antibodies or anti-TL1A/anti-TNF-α bispecific molecules in 96-well plate at 37° C. overnight. Each well was supplemented with 50 μl of Steady-glo Luciferase testing solution (Promega). Plate was covered and incubated while shaking for 10 minutes. Luciferase activity was analyzed by microbeta reader. 
     Tables 14.1-14.3 show anti-TL1A quality control (QC) and activity data for anti-TL1A antibodies, hetero Ig bispecific antibodies, IgG-scFv bispecific antibodies, and Fab&#39;s with both human and cynomolgus monkey TL1A. 
     
       
         
           
               
             
               
                 TABLE 14.1 
               
             
            
               
                   
               
               
                 anti-TL1A activity of anti-TL1A antibodies and Fab fragments 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 huTL1A (0.2 nM) 
                 cyno TL1A (2 nM) 
               
               
                 mAb 
                 mAb/Fab 
                 NF-kB IC50 nM 
                 NF-kB IC50 nM 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 3C6 
                 mAb 
                 0.153 
                 &gt;100 
               
               
                   
                 Fab 
                 0.162 
                 &gt;100 
               
               
                 17E9 
                 mAb 
                 0.168 
                 &gt;100 
               
               
                   
                 Fab 
                 70.83 
                 &gt;100 
               
               
                 3B3S65A 
                 mAb 
                 0.083 
                 0.595 
               
               
                   
                 Fab 
                 0.178 
                 0.434 
               
               
                 3B3/7B3 
                 mAb 
                 0.037 
                 0.390 
               
               
                   
                 Fab 
                 0.112 
                 0.647 
               
               
                 5G4 
                 mAb 
                 0.058 
                 0.622 
               
               
                   
                 Fab 
                 2.838 
                 10.06 
               
               
                 2G11 
                 mAb 
                 0.223 
                 0.938 
               
               
                   
                 Fab 
                 0.551 
                 1.637 
               
               
                 9C8 
                 mAb 
                 0.131 
                 0.721 
               
               
                   
                 Fab 
                 2.434 
                 2.748 
               
               
                 23B3 
                 mAb 
                 0.089 
                 0.619 
               
               
                   
                 Fab 
                 2.992 
                 2.629 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14.2 
               
             
            
               
                   
               
               
                 QC and activ1ty of hetero Ig bispecific antibodies 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                   
                 Activ1ty 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 Final 
                 Final 
                 SE-HPLC 
                 Caliper 
                 TL1A 
                 TNF-α 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 iPS No. 
                 TL1A 
                 TNF-α 
                 Conc. 
                 Yield 
                 HMW 
                 Main 
                 NR 1 
                 NR 2 
                 HC1 
                 H2 
                 LC1 
                 LC2 
                 (pM) 
                 (pM) 
               
               
                   
               
               
                 376541 
                 Certolizumab 
                 3B3 
                 12.05 
                 295 
                 0.7 
                  99.3 
                 72 
                 28 
                  42 
                 58 
                 51 
                 49 
                 187 
                  51 
               
               
                   
                 (parent) 
                 (variant) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 349461 
                 Certolizumab 
                 2G11 
                 10.49 
                 188 
                 0.7 
                  99.3 
                 45 
                 55 
                 100 
                 0 
                 51 
                 49 
                 1284 
                  60 
               
               
                   
                 (variant) 
                 (variant) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 349463 
                 Certolizumab 
                 23B3 
                 10.61 
                 450 
                 0.6 
                  99.4 
                 63 
                 37 
                  50 
                 50 
                 33 
                 67 
                 3464 
                  64 
               
               
                   
                 (variant) 
                 (VH4) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376542 
                 C234 
                 3B3 
                  9.76 
                 174 
                 0.0 
                 100.0 
                 71 
                 29 
                  44 
                 56 
                 50 
                 50 
                  147 
                 544 
               
               
                   
                   
                 (variant) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376543 
                 Certolizumab 
                 3B3 
                 13.60 
                 320 
                 0.4 
                  99.6 
                 75 
                 25 
                  43 
                 57 
                 51 
                 49 
                  158 
                 107 
               
               
                   
                 (variant) 
                 (variant) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14.3 
               
             
            
               
                   
               
               
                 QC and activ1ty of IgG-scFv bispecific antibodies 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Activ1ty 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                 Final 
                 Final 
                 SE-HPLC 
                 Caliper 
                 TL1A 
                 TNF-α 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 iPS 
                 IgG 
                 scFv 
                 Conc. 
                 Yield 
                 HMW 
                 Main 
                 Shoulder 
                 NR 1 
                 NR 2 
                 HC1 
                 LC1 
                 (pM) 
                 (pM) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 371213 
                 TNF-α 
                 TL1A 
                 12.40 
                 310 
                 .05 
                 98.3 
                 1.4 
                 11 
                 89 
                 81 
                 19 
                 61 
                 64 
               
               
                   
                 (adalimumab) 
                 (3B3 Var2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 371217 
                 TNF-α (C234) 
                 TL1A 
                 13.06 
                 290 
                 0.4 
                 98.5 
                 1.2 
                 39 
                 61 
                 81 
                 18 
                 69 
                 40 
               
               
                   
                   
                 (3B3 Var2/CC) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 369989 
                 TNF-α 
                 TL1A (9C8) 
                 13.83 
                 199 
                 0.3 
                 98.1 
                 1.1 
                 27 
                 73 
                 76 
                 18 
                 140 
                 59 
               
               
                   
                 (adalimumab) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 369995 
                 TNF-α (3.2) 
                 TL1A (9C8) 
                 15.81 
                 217 
                 1.0 
                 97.9 
                 1.6 
                   
                 100 
                 80 
                 17 
                 199 
                 53 
               
               
                 370001 
                 TNF-α (C234) 
                 TL1A (9C8) 
                 12.41 
                 289 
                 0.2 
                 98.2 
                 1.1 
                 42 
                 58 
                 84 
                 16 
                 119 
                 52 
               
               
                 369992 
                 TNF-α 
                 TL1A (9C8/CC) 
                 11.48 
                 169 
                 0.3 
                 98.2 
                 1.6 
                 16 
                 84 
                 84 
                 16 
                 151 
                 72 
               
               
                   
                 (adalimumab) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 370004 
                 TNF-α (C234) 
                 TL1A (9C8/CC) 
                 11.34 
                 111 
                 0.3 
                 98.2 
                 1.5 
                 45 
                 55 
                 84 
                 16 
                 163 
                 70 
               
               
                 370013 
                 TL1A (9C8) 
                 TNF-α (3.2) 
                 16.53 
                 464 
                 0.3 
                 98.3 
                 1.5 
                 41 
                 59 
                 79 
                 18 
                 171 
                 25 
               
               
                 371219 
                 TL1A 
                 TNF-α (3.2) 
                 10.9 
                 197 
                 0.4 
                 98.1 
                 1.4 
                   
                 100 
                 70 
                 20 
                 53 
                 22 
               
               
                   
                 (3B3 Var2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 371222 
                 TL1A 
                 TNF-α (3.2/CC) 
                 20.07 
                 157 
                 0.2 
                 98.3 
                 1.5 
                   
                 100 
                 78 
                 18 
                 86 
                 26 
               
               
                   
                 (3B3 Var2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 370018 
                 TL1A (23B3) 
                 TNF-α 
                 11.59 
                 112 
                 2.2 
                 96.5 
                 1.5 
                   
                 100 
                 86 
                 14 
                 157 
                 25 
               
               
                   
                   
                 (adalimumab) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 370021 
                 TL1A (23B3) 
                 TNF-α 
                 15.29 
                 321 
                 0.0 
                 100.0 
                 1.3 
                 7 
                 93 
                 86 
                 14 
                 139 
                 24 
               
               
                   
                   
                 (adalimumab/CC) 
               
               
                   
               
            
           
         
       
     
     Example 15 
     TNF-α Binding Assay 
     Goat anti-human Fc was first immobilized on SCMS sensor chip using amine coupling. Anti-TNF monoclonal antibodies, Fab fragment or TL1A/TNF bispecifics were injected to the chip at 10 μl/min for 1 min. Various concentration of human or cynomologus TNF protein from 0.78 to 25 nM in sample buffer (PBS, 0.005% P20, 0.1 mg/ml BSA) were injected at flow rate of 50 μl/min for 3 min association, 10 min dissociation. On rate, off rate and equilibrium dissociation constant were calculated using 1:1 binding model on BlAevaluation software. 
     Tables 15.1-15.3 show TNF-α binding activity data for anti-TNF-α antibodies, hetero Ig bispecific antibodies, IgG-scFv bispecific antibodies, and IgG-Fab bispecific antibodies. 
     
       
         
           
               
             
               
                 TABLE 15.1 
               
             
            
               
                   
               
               
                 TNF- α binding activ1ty of anti-TNF-α antibodies and Fab fragment 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 Human TNF-α 
                 Cyno TNF-α 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                 k a  (1/Ms) 
                 k d  (1/s) 
                 KD (M) 
                 k a  (1/MS) 
                 k d  (1/s) 
                 KD (M) 
               
               
                   
               
               
                 3.2 
                 mAb 
                 1.5E+06 
                  7.3E−05 
                 4.8E−11 
                 9.0E+05 
                  9.9E−05 
                 1.1E−10 
               
               
                   
                 Fab 
                   
                 &lt;5.0E−5  
                   
                   
                 &lt;5.0E−5  
                   
               
               
                 4.14 
                 mAb 
                 5.3E+05 
                  6.1E−05 
                 1.2E−10 
                 3.5E+05 
                  1.0E−04 
                 2.9E−10 
               
               
                   
                 Fab 
                   
                 &lt;5.0E−5  
                   
                   
                 &lt;5.0E−5  
                   
               
               
                 234 
                 mAb 
                 1.5E+06 
                  7.9E−05 
                 5.2E−11 
                 1.2E+06 
                  1.0E−04 
                 8.7E−11 
               
               
                   
                 Fab 
                   
                 &lt;5.0E−5  
                   
                   
                 &lt;5.0E−5  
                   
               
               
                 Adalimumab 
                 mAb 
                 7.1E+05 
                  6.1E−05 
                 8.6E−11 
                 6.4E+05 
                  5.5E−05 
                 8.7E−11 
               
               
                   
                 Fab 
                 4.5E+05 
                  1.4E−04 
                 3.2E−10 
                 3.6E+05 
                  8.1E−05 
                 2.3E−10 
               
               
                 Certolizumab 
                 Peg- 
                 3.9E+06 
                  6.2E−05 
                 1.6E−11 
                 6.4E+06 
                  2.2E−03 
                 3.5E−10 
               
               
                 pegol 
                 Fab 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15.2 
               
             
            
               
                   
               
               
                 TNF-α binding activ1ty of hetero Ig bispecific antibodies 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 iPS 
                 Lot 
                 anti-TNF-α 
                 anti-TL1A 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
                   
               
               
                   
               
               
                   
                 376541 
                 PL-42786 
                 Certolizumab 
                 3B3 variant 
                 3.2E+06 
                 4.4E−05* 
                 1.4E−11 
                   
               
               
                   
                   
                   
                 parent 
                 (322520) 
                   
                   
                   
                   
               
               
                   
                 376542 
                 PL-42789 
                 C234 
                 3B3 variant 
                 2.3E+06 
                 1.1E−04  
                 4.8E−11 
                   
               
               
                   
                   
                   
                   
                 (322520) 
                   
                   
                   
                   
               
               
                   
                 376543 
                 PL-42790 
                 Certolizumab 
                 3B3 variant 
                 3.6E+06 
                 5.5E−05  
                 1.6E−11 
                   
               
               
                   
                   
                   
                 variant 
                 (322520) 
                   
                   
                   
                   
               
               
                   
                 349461 
                 PL-42787 
                 Certolizumab 
                 2G11 Variant 
                 3.4E+06 
                 6.4E−05  
                 1.9E−11 
                   
               
               
                   
                   
                   
                 variant 
                   
                   
                   
                   
                   
               
               
                   
                 349463 
                 PL-42788 
                 Certolizumab 
                 23B3 VH4 
                 3.4E+06 
                 5.5E−05  
                 1.6E−11 
                   
               
               
                   
                   
                   
                 variant 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15.3 
               
             
            
               
                   
               
               
                 TNF-α binding activ1ty of IgG-scFv bispecific antibodies 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 iPS 
                 Lot 
                 IgG 
                 scFV 
                 ka (1/Ms) 
                 kd (1/s) 
                 KD (M) 
               
               
                   
               
               
                 369989 
                 PL-42765 
                 TNF-α 
                 TL1A (9C8) 
                 1.2E+06 
                 7.6E−05 
                 6.6E−11 
               
               
                   
                   
                 (Adalimumab) 
                   
                   
                   
                   
               
               
                 369992 
                 PL-42770 
                 TNF-α 
                 TL1A (9C8/CC) 
                 1.2E+06 
                 8.4E−05 
                 7.3E−11 
               
               
                   
                   
                 (Adalimumab) 
                   
                   
                   
                   
               
               
                 369995 
                 PL-42779 
                 TNF-α (3.2) 
                 TL1A (9C8) 
                 1.7E+06 
                 7.7E−05 
                 4.4E−11 
               
               
                 370001 
                 PL-42767 
                 TNF-α (C234) 
                 TL1A (9C8) 
                 1.7E+06 
                 9.9E−05 
                 5.9E−11 
               
               
                 370004 
                 PL-42785 
                 TNF-α (C234) 
                 TL1A (9C8/CC) 
                 1.7E+06 
                 1.0E−04 
                 5.9E−11 
               
               
                 370013 
                 PL-42781 
                 TL1A (9C8) 
                 TNF-α (3.2) 
                 1.3E+06 
                 8.8E−05 
                 6.5E−11 
               
               
                 370018 
                 PL-42763 
                 TL1A (23B3) 
                 TNF-α 
                 1.1E+06 
                 5.3E−05 
                 5.0E−11 
               
               
                   
                   
                   
                 (Adalimumab) 
                   
                   
                   
               
               
                 370021 
                 PL-42778 
                 TL1A (23B3) 
                 TNF-α 
                 1.1E+06 
                  4.9E−05* 
                 4.5E−11 
               
               
                   
                   
                   
                 (Adalimumab/CC) 
                   
                   
                   
               
               
                 371213 
                 PL-42771 
                 TNF-α 
                 TL1A 
                 1.3E+06 
                 8.2E−05 
                 6.5E−11 
               
               
                   
                   
                 (Adalimumab) 
                 (3B3.322520/CC) 
                   
                   
                   
               
               
                 371217 
                 PL-42782 
                 TNF-α (C234) 
                 TL1A 
                 1.9E+06 
                 8.8E−05 
                 4.8E−11 
               
               
                   
                   
                   
                 (3B3.322520/CC) 
                   
                   
                   
               
               
                 371219 
                 PL-42774 
                 TL1A 
                 TNF-α (3.2) 
                 1.3E+06 
                 5.5E−05 
                 4.2E−11 
               
               
                   
                   
                 (3B3.322520) 
                   
                   
                   
                   
               
               
                 371222 
                 PL-42783 
                 TL1A 
                 TNF-α (3.2/CC) 
                 1.3E+06 
                 6.3E−05 
                 4.7E−11 
               
               
                   
                   
                 (3B3.322520) 
               
               
                   
               
            
           
         
       
     
     Example 16 
     TNF-α Activity Assays 
     TNF-α activity assay was performed using TF-1 NF-kB reporter cell line. In brief, 1 ng/ml (EC90) of human or cynomolgus monkey TNF-α was incubated with 10 4  TF-1 NF-κB reporter cells in the presence of a serially diluted anti-TNF-α antibodies or TL1A/TNF-α bispecific molecules in 96-well plate at 37° C. overnight. 50 μl of Steady-glo Luciferase testing solution (Promega) was added to each well. Plate was covered and incubated while shaking for 10 minutes. Luciferase activity was analyzed by microbeta reader. 
     Table 16.1 shows anti-TNF-α QC and activity data for anti-TNF antibodies. Tables 14.2 and 14.3 show anti-TNF-α binding activity for hetero Ig bispecific antigen binding proteins, and IgG-scFv bispecific antigen binding proteins, respectively. 
     
       
         
           
               
             
               
                 TABLE 16.1 
               
             
            
               
                   
               
               
                 Anti-TNFα activity of TNF-α antibodies and Fab fragments 
               
            
           
           
               
               
               
               
            
               
                 Antibody 
                   
                 Human TNF (20 pM) 
                 Cyno TNF (20 pM) 
               
               
                 designation 
                 Modality 
                 IC50 (pM) NFkB 
                 IC50 (pM) NFkB 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 3.2 
                 mAb 
                 75 
                 334 
               
               
                   
                 Fab 
                 367 
                 2950 
               
               
                 4.14 
                 mAb 
                 127 
                 739 
               
               
                   
                 Fab 
                 526 
                 3105 
               
               
                 234 
                 mAb 
                 74 
                 138 
               
               
                   
                 Fab 
                 825 
                 2027 
               
               
                 adalimumab 
                 mAb 
                 119 
                 180 
               
               
                   
                 Fab 
                 670 
                 1800 
               
               
                 certolizurnab 
                 Peg-Fab 
                 91 
                 1.38 μM 
               
               
                 pegol 
               
               
                   
               
            
           
         
       
     
     Example 17 
     Bispecific Molecule Activity 
     The human and cynomolgus monkey TL1A and TNF-α binding activities of bispecific antigen binding proteins were determined as described in Examples 13 and 15. Data are shown in Table 17.1 
     
       
         
           
               
             
               
                 TABLE 17.1 
               
             
            
               
                   
               
               
                 human and cyno TL1A and TNF binding activ1ties 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 TL1A binding 
                 TNF binding  
               
               
                   
                 TL1A 
                 TNF-α 
                   
                 (Kd pM) 
                 (Kd pM) 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 leads 
                 warhead 
                 warhead 
                 iPS no. 
                 Hu TL1A 
                 Cyno TL1A 
                 Hu TNF 
                 Cyno TNF 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Hetero-Ig 
                 3B3 V2 
                 certolizumab 
                 376543 
                 11 
                 26 
                 12 
                 210 
               
               
                 IgG-scFy 
                 9C8/CC 
                 adalimumab 
                 381505 
                 120 
                 150 
                 21 
                 33 
               
               
                 IgG-scFy 
                 9C8/CC 
                 C234 
                 381489 
                 160 
                 94 
                 10 
                 19 
               
               
                 IgG-scFy 
                 3B3 V2/CC 
                 adalimumab 
                 381513 
                 1.8 
                 6.6 
                 19 
                 23 
               
               
                   
               
            
           
         
       
     
     The human and cynomolgus monkey TL1A and TNF-α blocking activities of bispecific antigen binding proteins were determined as described in Examples 14 and 16. Data are shown in Table 17.2. 
     
       
         
           
               
             
               
                 TABLE 17.2 
               
             
            
               
                   
               
               
                 human and cyno TL1A and TNF-α blocking activ1tes 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 TL1A assay IC50 pM 
                 TNF assay IC50 pM 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Anti-TNF-α 
                 iPS No. 
                 soluble 
                 mem  
                 soluble 
                 soluble 
                 mem  
                 soluble  
               
               
                 leads 
                 Anti-TL1A 
                 warhead 
                 warhead 
                 hu 
                 hu 
                 cyno 
                 hu 
                 hu 
                 cyno 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Hetero-Ig 
                 3B3 V2 
                 certolizumab 
                 376543 
                 45.5 
                 50.1 
                 49.7 
                 101.1 
                 942.6 
                 0.9  
                 uM 
               
               
                 IgG-scFy 
                 9C8/CC 
                 adalimumab 
                 381505 
                 52.2 
                 46.5 
                 45.3 
                 72.9 
                 719.5 
                 96.4 
                   
               
               
                 IgG-scFy 
                 9C8/CC 
                 C234 
                 381489 
                 66.7 
                 46.3 
                 62.6 
                 95.8 
                 639.2 
                 157.2 
                   
               
               
                 IgG-scFy 
                 3B3 V2/CC 
                 adalimumab 
                 381513 
                 35.6 
                 19.2 
                 23.7 
                 69.7 
                 906.3 
                 91.3 
               
               
                   
               
            
           
         
       
     
     Example 18 
     SNP Genotyping 
     To evaluate potential TL1A genotype association with expression, genomic DNA (gDNA) was isolated from healthy PBMC donors using Gentra Puregene Tissue kit from Qiagen. Genomic DNA were genotyped using TaqMan SNP genotyping assays for rs7848647, rs6478109, rs6478108, and rs3810936 assays from LifeTech and standard protocols on the Bio-Rad droplet digital PCR platform. Briefly, 10 ng of gDNA from each donor was mixed with ddPCR™ Supermix for Probes (Bio-Rad) and TaqMan SNP genotyping assay (LifeTech) for each SNP of interest. Droplets were generated for each reaction using the QX100™ droplet generator (Bio-Rad) and subjected to thermal cycling on a C1000 Touch™ thermal cycler (Bio-Rad). Following amplification, droplet fluorescence was read on a QX100™ droplet reader (Bio-Rad) and data were analyzed using QuantaSoft software (Bio-Rad). Donors were considered homozygous risk haplotype if only risk alleles were present at all 4 genotyped SNPs (rs7848647, rs6478109, rs6478108, and rs3810936). Donors were considered homozygous non-risk haplotype if only non-risk alleles were present at all 4 genotyped SNPs. Donors were considered heterozygous haplotype if both risk and non-risk alleles were present at all 4 genotyped SNPs. Donors that were considered “recombinant” had only homozygous risk alleles at rs7848647, rs6478109, and rs6478108, but were heterozygous (risk and non-risk alleles present) at rs3810936. 
     The presence of synonymous SNP (rs3810936) in the exon 4 of the TL1A gene enabled us to track risk allele vs non-risk allele expression by droplet digit PCR (ddPCR) using allelic specific fluorescent probes in the allelic expression imbalance (AEI) study. The frequency of risk vs non-risk allele usage in heterozygous PBMC at basal level or after immune complex stimulation was evaluated at various time points. In brief, PBMCs heterozygous for TL1A genotype were treated with immune complex for various lengths of time. RNA was then isolated from PBMC using an RNeasy mini kit with on-column DNase I digestion from Qiagen. RNA from each sample was converted to cDNA using the High Capacity cDNA Reverse Transcription Kit according to manufacturer&#39;s instructions (LifeTech). Allele-specific expression at rs3810936 was determined using TaqMan SNP genotyping assay specific for rs3810936 (LifeTech) and standard protocols on the Bio-Rad droplet digital PCR platform. In brief, the amount of input cDNA was first optimized for TNFSF15 expression at each time point in order to increase precision by maximizing the number of positive droplets counted for each sample by QuantaSoft software. The optimal amount of cDNA per sample was mixed with ddPCR™ Supermix for Probes (Bio-Rad) and TaqMan SNP genotyping assay (LifeTech) for rs3810936. Droplets were generated for each reaction using the QX100′ droplet generator (Bio-Rad) and subjected to thermal cycling on a C1000 Touch™ thermal cycler (Bio-Rad). Following amplification, droplet fluorescence was read on a QX100 droplet reader (Bio-Rad). Data were analyzed using QuantaSoft software (Bio-Rad) and copies/ml of each allele at rs3810936 was determined. The allelic expression ratio was calculated by dividing the copies/ml of the risk allele by the copies/ml of the non-risk allele. Total copy number of each allele was normalized by the input amount of cDNA (copies/ng), and the fold-induction for each allele at each time point was calculated by dividing the copies/ng at the time point of interest by the copies/ng at baseline (0 hr). Higher induction of TL1A risk allele was observed in PBMC after immune complex treatment compared to non-risk allele in heterozygous PBMC in allelic expression imbalance study. 
     The inventor(s) identified a small number of donors homozygous for risk alleles at rs7848647, rs6478109, and rs6478108, but heterozygous at rs3810936, likely due to recombination between rs6478109 and synonymous SNP rs3810936. Since these donors are heterozygous at synonymous SNP rs381093, the inventor(s) could track the risk allele and non-risk allele usage and evaluate if the allelic expression imbalance was still retained in these donors. Interestingly, compared to heterozygous donors, no allelic expression imbalance was detected in these recombinant individuals before or after immune complex stimulation. This demonstrates that synonymous SNP rs381093 itself is not responsible for allelic imbalance regulation. The regulatory SNPs likely derive from rs7848647, rs6478109, rs6478108 and/or other SNPs 5′ to the synonymous SNP rs3810936. 
     To evaluate if TL1A allelic expression imbalance data correlate with expression quantitative trait loci (eQTL), PBMC donors with homozygous TL1A risk allele, homozygous non-risk alleles or heterozygous alleles were treated with immune complex. RNA from each sample was isolated and then converted to cDNA as described previously. Total expression of TL1A (TNFSF15) in each donor was determined by digital PCR. In brief, cDNA from each sample was mixed with ddPCR™ Supermix for Probes (Bio-Rad) and PrimeTime® qPCR assay ID Hs.PT.56a.41003970. Droplets were generated for each reaction using the QX100™ droplet generator (Bio-Rad) and subjected to thermal cycling on a C1000 Touch™ thermal cycler (Bio-Rad). Following amplification, droplet fluorescence was read on a QX100™ droplet reader (Bio-Rad). Data were analyzed using QuantaSoft software (Bio-Rad) and copies/μl of TL1A was determined and normalized for the amount of input cDNA (copies/ng). P values for differences in TL1A expression levels between different TNFSF15 haplotypes were determined using the student t test. TL1A is expressed at a very low basal level in PBMCs, but strongly induced after immune complex stimulation. At basal level, PBMCs from donors homozygous for TL1A risk SNPs have lower TL1A mRNA compared to non-risk homozygous donors, albeit at very low copy number, consistent with the allelic expression imbalance study which demonstrated lower ratio of risk allele versus non-risk allele without stimulation. However, after immune complex stimulation, PBMCs from donors homozygous for TL1A risk SNPs have higher TL1A expression compared to non-risk homozygous donors. These data, together, demonstrated TL1A risk SNPs regulate higher TL1A induction. The inventor(s) speculate that in inflammatory conditions such as IBD, donors carrying TL1A risk SNPs likely encounter higher TL1A induction upon encounter of stimuli such as cytokines, opsonized or non-opsonized microbes. Therefore, TL1A risk SNPs could be used for patient stratification for TL1A inhibitor or anti-TL1A/anti-TNF-α bispecific inhibitors. 
     To evaluate if TL1A genotype-mediated expression regulation is tissue-specific, HUVEC cells from various donors were genotyped as described previously. Genotyped HUVEC cells from various donors were treated with IL-1 at various time points. Total copy number of each allele was measured as described previously. The allelic expression ratio was calculated by dividing the copies/ml of the risk allele by the copies/ml of the non-risk allele. No allelic expression imbalance was observed in HUVEC cells with or without IL-1 treatment. 
     Example 19 
     Mouse IBD Models 
     To evaluate role of TL1A in IBD, a series of preclinical experiments were performed in mouse IBD models. The effect of TL1A inhibition was evaluated in mouse mdr1a−/− spontaneous colitis model. Mdr1a gene encodes multiple drug resistance gene for P-glycoprotein 170. Mice with knockout of mdr1a gene are prone to develop spontaneous colitis from 12 weeks of age due to weakened intestinal barrier. Disease course is affected by gut microbes. In our experiment, female Mdr1a −/−  or wild type FVB controls were obtained from Taconic at 4-6 weeks of age. Animal weight and clinical disease activity was monitored regularly. The clinical disease activity score was arrived at using a summation of the scores obtained evaluating anal inflammation (0=none, 1=mild, 2=moderate, 3=severe, 4=rectal prolapse) and stool consistency (0=normal, 1=moist/sticky, 2=soft, 3=diarrhea, 4=bloody). Mice (n=10, 6-7 weeks age) were randomized to different groups based upon baseline clinical disease activity measurements, and treated intra-peritoneally once a week with 500 μg of anti-mouse TL1A antibody, anti-IL23p19 antibody, anti-mouse IL17RA antibody, or mouse isotype control or no treatment once per week for 8 weeks. Mice were then sacrificed, sections of the intestine were taken and processed for H&amp;E staining prior to scoring for disease by a pathologist. The following score was assigned based upon the histopathology: 0=normal; 1=minimal, mononuclear infiltrate and/or epithelial hypertrophy/hyperplasia; 2=mild, mononuclear infiltrate and/or epithelial hypertrophy/hyperplasia; 3=moderate, mononuclear infiltrate and/or epithelial hypertrophy/hyperplasia with rare crypt abscesses; 4=marked, same as for 3 plus abundant crypt abscesses and crypt dropout and/or focal ulceration; 5=severe, same as for 4 but with large areas of crypt dropout and/or extensive areas of ulceration. Statistical analysis was performed using one-way ANOVA with Dunett&#39;s compared to mIgG1 group. In conclusion, prophylactic treatment with anti-TL1A mAb inhibited spontaneous colitis development in mdr1a −/−  Mice. 
     The inventor(s) also evaluated if challenge with TL1A protein will exacerbate colitis development in mdr1a −/−  mice, which are prone to develop spontaneous colitis from 12 weeks of age due to a weakened intestinal barrier. Mdr1a −/−  mice or wild-type control mice at 6-8 weeks of age were treated intra-peritoneally once a week with 150 μg of recombinant mFc-TL1A fusion protein or isotype control three times each week for 4 weeks. Clinical disease activity was monitored by evaluating anal inflammation and stool consistency as described previously. After 4 weeks of treatment, mice were sacrificed, sections of the intestine were taken and processed for H&amp;E staining prior to scoring for disease as described previously. Statistical analysis was performed using one-way ANOVA with Dunett&#39;s compared to mIgG1 group. Challenge of TL1A protein severely exacerbated colitis development in mdr1a −/−  mice but not in wild-type mice. 
     The inventor(s) further examined the impact of TL1A challenge in lamina propria lymphocytes in mdr1a −/−  mice versus wild-type mice. In brief, lamina propria lymphocytes were isolated as previously described (D&#39;Souza W N et al., JI, V168:5566-5572, 2002). In brief, intestines were isolated, cut open longitudinally, rinsed with buffer and cut into 0.5 cm pieces. They were then washed twice in EDTA and the supernatants were discarded. The pieces were washed with RPMI and incubated in collagenase/DNase for 30 minutes. The isolated cells were run through a percoll gradient and the cells collected at the interphase were stained for surface antigens and analyzed by FACS. TL1A challenge in mdr1a −/−  mice resulted in increased inflammatory cells in lamina propria. 
     Example 20 
     Pharmacodynamics Studies 
     To evaluate if TL1A and TNF challenges result in similar or different pharmacodynamics effects, C57Bl/6 mice (8 week, female) were first intraperitonially injected with 500 μg/mice of anti-mouse TL1A, anti-mouse TNF-α or PBS. After 4 hours, the mice were then challenged with 100 μg/mice of TL1A or 10 μg/mice of TNF-α or without challenging. The sera were collected after 24 hours. The cytokines were measured by MSD (IL-22 was measured by ELISA). Challenge with TL1A or TNF protein in mice resulted in distinct cytokine induction. TL1A challenge mainly induced IFN-γ and IL-5, whereas TNF challenge induced IL-6, IL-8 and IL-10. 
     The inventor(s) evaluated cytokine induction by TL1A or TNF treatment in human PBMC. In brief, PBMC freshly isolated from human blood were cultured in media (RPMI1640 supplemented with 10% FBS, 2 mM glutamine, 1 mM sodium pyruvate, 5×10 −5  M 2-ME, and antibiotics) in the presence of 100 ng/ml of human TL1A or TNF-α. Supernatant was collected after 72 hours. The cytokines in the supernatant were measured by MSD (IL-22 was measured by ELISA). Similar to the mice challenge experiment, treatment of TL1A with human PBMC resulted in induction of IFN-γ, IL-5 and IL-22, whereas TNF treatment resulted in increase of IL-8, IL-10 and MCP-1. Therefore, TL1A and TNF induce distinct cytokine profiles in human PBMC. 
     Example 21 
     IgG-Fab Molecules 
     Bispecific antigen binding proteins were prepared with a subset of the anti-TNFα and anti-TL1A antibodies. In some embodiments of this IgG-Fab format, a polypeptide comprising a VH-CH1 domain from a second antibody is fused through a peptide linker to the carboxyl-terminus of the heavy chain of a first antibody to form a modified heavy chain. A polypeptide comprising the remaining domains of the Fab fragment from the first antibody (i.e. a VL-CL domain) is co-expressed with the light chain of the first antibody and the modified heavy chain to produce the complete molecule. Assembly of the full molecule creates a tetravalent binding protein having two antigen binding domains against a first antigen located on the amino terminal side of a dimerized immunoglobulin Fc region and two antigen binding domains against a second antigen located on the carboxyl terminal side of the dimerized Fc region. 
     The TNFα/TL1A IgG-Fab consists of two antigen binding domains, one directed against TNFα and the other against TL1A. The DNA molecules encoding TNFα/TL1A IgG-Fab molecules contain fragments encoding an anti-TNFα (or anti-TL1A) antibody light chain, an anti-TNFα (or anti-TL1A) antibody heavy chain in which the C-terminus is fused to (i) an anti-TL1A (or anti-TNFα) antibody light chain or (ii) an anti-TL1A (or anti-TNFα) Fd (VH-CH1), and a third polypeptide comprising the other half of the Fab fragment to complete the carboxy-terminal binding domain; for example, (i) an anti-TL1A (or anti-TNFα) Fd or (ii) an anti-TL1A (or anti-TNFα) antibody light chain. The IgG-Fab bispecific molecules contain charge pair mutations introduced into CH1 and CL domains of each Fab region (Fab 1 and Fab 2 as illustrated in  FIG. 3 ). The charge pairs are designed to allow preferential assembly of anti-TNFAR light chain/VHCH1(Fd) pair and anti-TL1A light chainNHCH1 (Fd) pair. As an additional approach to promote correct pairing of the light chain/VHCH1 (Fd) pair, for a subset of the IgG-Fab molecules generated, the CL and CH1 regions in the carboxyl-terminal Fab (i.e. Fab 2) were swapped such that the polypeptide fused to the carboxyl-terminal region of the heavy chain of the second antibody comprised VL and CH1 regions from the first antibody and the second polypeptide comprised VH and CL regions from the first antibody. See molecules listed in Tables 21.1 and 21.3, with the VL and VH CDRs listed in Table 21.2A and 21.2B and purity listed in Table 21.4. The DNA molecules were generated by synthesized gBlocks and cloned into the pTT5.1 vector. These expression vectors were used to transfect and express the TNFα/TL1A bispecific molecules in human 293-6E cells. 144 different IgG-Fab bispecific molecules were generated. The full sequences for each molecule are set forth in Table 21.1 and the Sequence Listing. 
     The IgG-Fab molecules were purified using affinity captured by MabSelect SuRe chromatography (GE Life Sciences, Piscataway, N.J.) using a Large Format Autosampler (LFAS, Amgen, Inc., Thousand Oaks, Calif.). Clarified, conditioned media was loaded onto a 1 mL HiTrap MabSelect SuRe column (GE Life Sciences, Piscataway, N.J.) equilibrated with Dulbecco&#39;s phosphate buffered saline without divalent cations (D-PBS, Life Technologies, Grand Island, N.Y.). MabSelect columns were washed with 8 column volumes of D-PBS and eluted with 100 mM acetic acid, pH 3.6. When protein A eluates had an absorbance above 5 mAU at 280 nm, the eluent was directly loaded onto a HiTrap Desalting column (GE Life Sciences, Piscataway, N.J.) and developed with 1.2 column volumes of 10 mM sodium acetate, 150 mM NaCl, pH 5.0. When desalting eluates had an absorbance above 3 mAU at 280 nm, sample collection was triggered and fractions were collected in 96-well deepwell blocks to a maximum of 2 mL each. Sample purity was determined by Caliper LabChip analysis under reducing (with 2% 2-mercaptoethanol) and non-reducing (with 25 mM iodoacetamide) conditions. Analytical SEC was carried out using a Zenix-C SEC-300 column (Sepax Technologies, Newark, Del.) with an isocratic elution in 50 mM sodium phosphate, 250 mM NaCl, pH 6.9 over 8′. 
     The IgG-Fab molecules were tested for their expressability (titer and recovery) and activity. The results are shown in  FIGS. 27 to 33  and in Table 21.3. In Table 21.3 and throughout, “ada” refers to adalimumab. 
     TL1A activity assay was performed using TF-1 NF-κB reporter cell line. In brief, 30 ng/ml (EC90) of human or cynomolgus monkey TL1A was incubated with 10 4  TF-1 NF-κB reporter cells in the presence of serially diluted anti-TL1A antibodies or TL1A/TNF-α bispecific molecules in 96-well plate at 37° C. overnight. Each well was supplemented with 50 μl of Steady-glo Luciferase testing solution (Promega). Plate was covered and incubated while shaking for 10 minutes. Luciferase activity was analyzed by microbeta reader. 
     TNFα activity assay was performed using TF-1 NF-kB reporter cell line. In brief, 1 ng/ml (EC90) of human or cynomolgus monkey TNF-α was incubated with 10 4  TF-1 NF-κB reporter cells in the presence of a serially diluted anti-TNF-α antibodies or TL1A/TNFα bispecific molecules in 96-well plate at 37° C. overnight. 50 μl of Steady-glo Luciferase testing solution (Promega) was added to each well. Plate was covered and incubated while shaking for 10 minutes. Luciferase activity was analyzed by microbeta reader. 
     
       
         
           
               
             
               
                 TABLE 21.1 
               
             
            
               
                   
               
               
                 anti-TL1A anti-TNF-α IgG-Fab molecule amino acid sequences 
               
            
           
           
               
               
               
               
               
            
               
                 Molecule 
                   
                   
                   
                   
               
               
                 Designation 
                   
                 Light chain 1 
                 Light chain 2 
                 Heavy chain 
               
               
                 (iPS no.) 
                 IgG source, Fab source, aa substitutions 
                 SEQ ID NO 
                 SEQ ID NO 
                 SEQ ID NO 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 376597 
                 001_Adalimumab_IgG.001_3B3v2_VH_CH1 
                 1254 
                 1256 
                 1258 
               
               
                   
                 (S183E) 
                   
                   
                   
               
               
                 376601 
                 002_Adalimumab_IgG.001_9C8_VH_CH1(S183E) 
                 1260 
                 1262 
                 1264 
               
               
                 376605 
                 003_Adalimumab_IgG.001_23B3_VH4_VH_ 
                 1266 
                 1268 
                 1270 
               
               
                   
                 CH1(S183E) 
                   
                   
                   
               
               
                 376609 
                 004_3.2_IgG.001_3B3v2_VH_CH1(S183E) 
                 1272 
                 1274 
                 1276 
               
               
                 376613 
                 005_3.2_IgG.001_9C8_VH_CH1(S183E) 
                 1278 
                 1280 
                 1282 
               
               
                 376617 
                 006_3.2_IgG.001_23B3_VH4_VH_CH1(S183E) 
                 1284 
                 1286 
                 1288 
               
               
                 376621 
                 007_3B3v2_IgG.001_Adalimumab_VH_CH1 
                 1290 
                 1292 
                 1294 
               
               
                   
                 (S183E) 
                   
                   
                   
               
               
                 376625 
                 008_3B3v2_IgG.001_3.2_VH_CH1(S183E) 
                 1296 
                 1298 
                 1300 
               
               
                 376629 
                 009_9C8_IgG.001_Adalimumab_VH_CH1(S183E) 
                 1302 
                 1304 
                 1306 
               
               
                 376633 
                 010_9C8_IgG.001_3.2_VH_CH1(S183E) 
                 1308 
                 1310 
                 1312 
               
               
                 376637 
                 011_23B3_VH4_IgG.001_Adalimumab_VH_ 
                 1314 
                 1316 
                 1318 
               
               
                   
                 CH1(S183E) 
                   
                   
                   
               
               
                 376641 
                 012_23B3_VH4_IgG.001_3.2_VH_CH1(S183E) 
                 1320 
                 1322 
                 1324 
               
               
                 376645 
                 013_Adalimumab_CK(S176K)_Fc_3B3v2_V 
                 1326 
                 1328 
                 1330 
               
               
                   
                 H_CH1(S183E) 
                   
                   
                   
               
               
                 376651 
                 014_Adalimumab_CK(S176K)_Fc_9C8_VH_ 
                 1332 
                 1334 
                 1336 
               
               
                   
                 CH1(S183E) 
                   
                   
                   
               
               
                 376655 
                 015_Adalimumab_CK(S176K)_Fc_23B3_VH4_ 
                 1338 
                 1340 
                 1342 
               
               
                   
                 VH_CH1(S183E) 
                   
                   
                   
               
               
                 376659 
                 016_3.2_CL(S176K)_Fc_3B3v2_VH_CH1(S 
                 1344 
                 1346 
                 1348 
               
               
                   
                 183E) 
                   
                   
                   
               
               
                 376665 
                 017_3.2_CL(S176K)_Fc_9C8_VH_CH1(S183E) 
                 1350 
                 1352 
                 1354 
               
               
                 376669 
                 018_3.2_CL(S176K)_Fc_23B3_VH4_VH_C 
                 1356 
                 1358 
                 1360 
               
               
                   
                 H1(S183E) 
                   
                   
                   
               
               
                 376673 
                 019_3B3v2_CK(S176K)_Fc_Adalimumab_V 
                 1362 
                 1364 
                 1366 
               
               
                   
                 H_CH1(S183E) 
                   
                   
                   
               
               
                 376679 
                 020_3B3v2_CK(S176K)_Fc_3.2_VH_CH1(S 
                 1368 
                 1370 
                 1372 
               
               
                   
                 183E) 
                   
                   
                   
               
               
                 376683 
                 021_9C8_CK(S176K)_Fc_Adalimumab_VH_ 
                 1374 
                 1376 
                 1378 
               
               
                   
                 CH1(S183E) 
                   
                   
                   
               
               
                 376689 
                 022_9C8_CK(S176K)_Fc_3.2_VH_CH1(S183E) 
                 1380 
                 1382 
                 1384 
               
               
                 376693 
                 023_23B3_VH4_CK(S176K)_Fc_Adalimumab_ 
                 1386 
                 1388 
                 1390 
               
               
                   
                 VH_CH1(S183E) 
                   
                   
                   
               
               
                 376699 
                 024_23B3_VH4_CK(S176K)_Fc_3.2_VH_C 
                 1392 
                 1394 
                 1396 
               
               
                   
                 H1(S183E) 
                   
                   
                   
               
               
                 376703 
                 025_Adalimumab_IgG.001_3B3v2_VH_CK 
                 1398 
                 1400 
                 1402 
               
               
                   
                 (S176E) NA 
                   
                   
                   
               
               
                 376709 
                 026_Adalimumab_IgG.001_9C8_VH_CK(S176E) 
                 1404 
                 1406 
                 1408 
               
               
                 376715 
                 027_Adalimumab_IgG.001_23B3_VH4_VH_ 
                 1410 
                 1412 
                 1414 
               
               
                   
                 CK(S176E) 
                   
                   
                   
               
               
                 376721 
                 028_3.2_IgG.001_3B3v2_VH_CK(S176E) 
                 1416 
                 1418 
                 1420 
               
               
                 376725 
                 029_3.2_IgG.001_9C8_VH_CK(S176E) 
                 1422 
                 1424 
                 1426 
               
               
                 376729 
                 030_3.2_IgG.001_23B3_VH4_VH_CK(S176E) 
                 1428 
                 1430 
                 1432 
               
               
                 376733 
                 031_3B3v2_IgG.001_Adalimumab_VH_CK 
                 1434 
                 1436 
                 1438 
               
               
                   
                 (S176E) 
                   
                   
                   
               
               
                 376739 
                 032_3B3v2_IgG.001_3.2_VH_CL(S176E) 
                 1440 
                 1442 
                 1444 
               
               
                 376745 
                 033_9C8_IgG.001_Adalimumab_VH_CK(S1 
                 1446 
                 1448 
                 1450 
               
               
                   
                 76E) 
                   
                   
                   
               
               
                 376750 
                 034_9C8_IgG.001_3.2_VH_CL(S176E) 
                 1452 
                 1454 
                 1456 
               
               
                 376755 
                 035_23B3_VH4_IgG.001_Adalimumab_VH_ 
                 1458 
                 1460 
                 1462 
               
               
                   
                 CK(S176E) 
                   
                   
                   
               
               
                 376760 
                 036_23B3_VH4_IgG.001_3.2_VH_CL(S176E) 
                 1464 
                 1466 
                 1468 
               
               
                 376765 
                 037_Adalimumab_CK(S176K)_Fc_3B3v2_V 
                 1470 
                 1472 
                 1474 
               
               
                   
                 H_CK(S176E) 
                   
                   
                   
               
               
                 376770 
                 038_Adalimumab_CK(S176K)_Fc_9C8_VH_ 
                 1476 
                 1478 
                 1480 
               
               
                   
                 CK(S176E) 
                   
                   
                   
               
               
                 376775 
                 039_Adalimumab_CK(S176K)_Fc_23B3_VH4_ 
                 1482 
                 1484 
                 1486 
               
               
                   
                 VH_CK(S176E) 
                   
                   
                   
               
               
                 376779 
                 040_3.2_CL(S176K)_Fc_3B3v2_VH_CK(S1 
                 1488 
                 1490 
                 1492 
               
               
                   
                 76E) 
                   
                   
                   
               
               
                 376784 
                 041_3.2_CL(S176K)_Fc_9C8_VH_CK(S176E) 
                 1494 
                 1496 
                 1498 
               
               
                 376789 
                 042_3.2_CL(S176K)_Fc_23B3_VH4_VH_C 
                 1500 
                 1502 
                 1504 
               
               
                   
                 K(S176E) 
                   
                   
                   
               
               
                 376793 
                 043_3B3v2_CK(S176K)_Fc_Adalimumab_V 
                 1506 
                 1508 
                 1510 
               
               
                   
                 H_CK(S176E) 
                   
                   
                   
               
               
                 376797 
                 044_3B3v2_CK(S176K)_Fc_3.2_VH_CL(S176E) 
                 1512 
                 1514 
                 1516 
               
               
                 376801 
                 045_9C8_CK(S176K)_Fc_Adalimunab_VH_ 
                 1518 
                 1520 
                 1522 
               
               
                   
                 CK(S176E) 
                   
                   
                   
               
               
                 376806 
                 046_9C8_CK(S176K)_Fc_3.2_VH_CL(S176E) 
                 1524 
                 1526 
                 1528 
               
               
                 376811 
                 047_23B3_VH4_CK(S176K)_Fc_Adalimumab_ 
                 1530 
                 1532 
                 1534 
               
               
                   
                 VH_CK(S176E) 
                   
                   
                   
               
               
                 376816 
                 048_23B3_VH4_CK(S176K)_Fc_3.2_VH_C 
                 1536 
                 1538 
                 1540 
               
               
                   
                 L(S176E) 
                   
                   
                   
               
               
                 376821 
                 059_Adalimumab_IgG.002_3B3v2_VH_CH1 
                 1542 
                 1544 
                 1546 
               
               
                   
                 (S183E).001 
                   
                   
                   
               
               
                 376826 
                 060_Adalimumab_IgG.002_9C8_VH_CH1(S 
                 1548 
                 1550 
                 1552 
               
               
                   
                 183E).001 
                   
                   
                   
               
               
                 376830 
                 061_Adalimumab_IgG.002_23B3_VH4_VH_ 
                 1554 
                 1556 
                 1558 
               
               
                   
                 CH1(S183E).001 
                   
                   
                   
               
               
                 376834 
                 062_3.2_IgG.002_3B3v2_VH_CH1(S183E).001 
                 1560 
                 1562 
                 1564 
               
               
                 376868 
                 063_3.2_IgG.002_9C8_VH_CH1(S183E).001 
                 1566 
                 1568 
                 1570 
               
               
                 376842 
                 064_3.2_IgG.002_23B3_VH4_VH_CH1(S18 
                 1572 
                 1574 
                 1576 
               
               
                   
                 3E).001 
                   
                   
                   
               
               
                 376846 
                 065_3B3v2_IgG.002_Adalimumab_VH_CH1 
                 1578 
                 1580 
                 1582 
               
               
                   
                 (S183E).001 
                   
                   
                   
               
               
                 376850 
                 066_3B3v2_IgG.002_3.2_VH_CH1(S183E).001 
                 1584 
                 1586 
                 1588 
               
               
                 376854 
                 067_9C8_IgG.002_Adalimumab_VH_CH1(S 
                 1590 
                 1592 
                 1594 
               
               
                   
                 183E).001 
                   
                   
                   
               
               
                 376858 
                 068_9C8_IgG.002_3.2_VH_CH1(S183E).001 
                 1596 
                 1598 
                 1600 
               
               
                 376862 
                 069_23B3_VH4_IgG.002_Adalimunab_VH_ 
                 1602 
                 1604 
                 1606 
               
               
                   
                 CH1(S183E).001 
                   
                   
                   
               
               
                 376867 
                 070_23B3_VH4_IgG.002_3.2_VH_CH1(S183E).001 
                 1608 
                 1610 
                 1612 
               
               
                 376872 
                 071_Adalimumab_CK(S176K)_Fc.001_3B3v2_ 
                 1614 
                 1616 
                 1618 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376879 
                 072_Adalimumab_CK(S176K)_Fc.001_9C8_ 
                 1620 
                 1622 
                 1624 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376884 
                 073_Adalimumab_CK(S176K)_Fc.001_23B3_ 
                 1626 
                 1628 
                 1630 
               
               
                   
                 VH4_VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376889 
                 074_3.2_CL(S176K)_Fc.001_3B3v2_VH_C 
                 1632 
                 1634 
                 1636 
               
               
                   
                 H1(S183E).001 
                   
                   
                   
               
               
                 376896 
                 075_3.2_CL(S176K)_Fc.001_9C8_VH_CH1 
                 1638 
                 1640 
                 1642 
               
               
                   
                 (S183E).001 
                   
                   
                   
               
               
                 376901 
                 076_3.2_CL(S176K)_Fc.001_23B3_VH4_V 
                 1644 
                 1646 
                 1648 
               
               
                   
                 H_CH1(S183E).001 
                   
                   
                   
               
               
                 376906 
                 077_3B3v2_CK(S176K)_Fc.001_Adalimumab_ 
                 1650 
                 1652 
                 1654 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376913 
                 078_3B3v2_CK(S176K)_Fc.001_3.2_VH_C 
                 1656 
                 1658 
                 1660 
               
               
                   
                 H1(S183E).001 
                   
                   
                   
               
               
                 376918 
                 079_9C8_CK(S176K)_Fc.001_Adalimumab_ 
                 1662 
                 1664 
                 1666 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376925 
                 080_9C8_CK(S176K)_Fc.001_3.2_VH_CH1 
                 1668 
                 1670 
                 1672 
               
               
                   
                 (S183E).001 
                   
                   
                   
               
               
                 376930 
                 081_23B3_VH4_CK(S176K)_Fc.001_Adalimumab_ 
                 1674 
                 1676 
                 1678 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376937 
                 082_23B3_VH4_CK(S176K)_Fc.001_3.2_ 
                 1680 
                 1682 
                 1684 
               
               
                   
                 VH_CH1(S183E).001 
                   
                   
                   
               
               
                 376941 
                 083_Adalimumab_IgG.002_3B3v2_VH_CK 
                 1686 
                 1688 
                 1690 
               
               
                   
                 (S176E).001 
                   
                   
                   
               
               
                 376948 
                 084_Adalimumab_IgG.002_9C8_VH_CK(S1 
                 1692 
                 1694 
                 1696 
               
               
                   
                 76E).001 
                   
                   
                   
               
               
                 376955 
                 085_Adalimumab_IgG.002_23B3_VH4_VH_ 
                 1698 
                 1700 
                 1702 
               
               
                   
                 CK(S176E).001 
                   
                   
                   
               
               
                 376962 
                 086_3.2_IgG.002_3B3v2_VH_CK(S176E).001 
                 1704 
                 1706 
                 1708 
               
               
                 376967 
                 087_3.2_IgG.002_9C8_VH_CK(S176E).001 
                 1710 
                 1712 
                 1714 
               
               
                 376972 
                 088_3.2_IgG.002_23B3_VH4_VH_CK(S176E).001 
                 1716 
                 1718 
                 1720 
               
               
                 376976 
                 089_3B3v2_IgG.002_Adalimumab_VH_CK 
                 1722 
                 1724 
                 1726 
               
               
                   
                 (S176E).001 
                   
                   
                   
               
               
                 376983 
                 090_3B3v2_IgG.002_3.2_VH_CL(S176E).001 
                 1728 
                 1730 
                 1732 
               
               
                 376990 
                 091_9C8_IgG.002_Adalimumab_VH_CK(S1 
                 1734 
                 1736 
                 1738 
               
               
                   
                 76E).001 
                   
                   
                   
               
               
                 376995 
                 092_9C8_IgG.002_3.2_VH_CL(S176E).001 
                 1740 
                 1742 
                 1744 
               
               
                 377000 
                 093_23B3_VH4_IgG.002_Adalimumab_V14 
                 1746 
                 1748 
                 1750 
               
               
                   
                 CK(S176E).001 
                   
                   
                   
               
               
                 377005 
                 094_23B3_VH4_IgG.002_3.2_VH_CL(S176E).001 
                 1752 
                 1754 
                 1756 
               
               
                 377010 
                 095_Adalimumab_CK(S176K)_Fc.001_3B3v 
                 1758 
                 1760 
                 1762 
               
               
                   
                 2_VH_CK(S176E).001 
                   
                   
                   
               
               
                 377015 
                 096_Adalimumab_CK(S176K)_Fc.001_9C8_ 
                 1764 
                 1766 
                 1768 
               
               
                   
                 VH_CK(S176E).001 
                   
                   
                   
               
               
                 377020 
                 097_Adalimumab_CK(S176K)_Fc.001_23B3_ 
                 1770 
                 1772 
                 1774 
               
               
                   
                 VH4_VH_CK(S176E).001 
                   
                   
                   
               
               
                 377025 
                 098_3.2_CL(S176K)_Fc.001_3B3v2_VH_C 
                 1776 
                 1778 
                 1780 
               
               
                   
                 K(S176E).001 
                   
                   
                   
               
               
                 377030 
                 099_3.2_CL(S176K)_Fc.001_9C8_VH_CK(S 
                 1782 
                 1784 
                 1786 
               
               
                   
                 176E).001 
                   
                   
                   
               
               
                 377035 
                 100_3.2_CL(S176K)_Fc.001_23B3_VH4_V 
                 1788 
                 1790 
                 1792 
               
               
                   
                 H_CK(S176E).001 
                   
                   
                   
               
               
                 377040 
                 101_3B3v2_CK(S176K)_Fc.001_Adalimumab_ 
                 1794 
                 1796 
                 1798 
               
               
                   
                 VH_CK(S176E).001 
                   
                   
                   
               
               
                 377045 
                 102_3B3v2_CK(S176K)_Fc.001_3.2_VH_C 
                 1800 
                 1802 
                 1804 
               
               
                   
                 L(S176E).001 
                   
                   
                   
               
               
                 377050 
                 103_9C8_CK(S176K)_Fc.001_Adalimumab_ 
                 1806 
                 1808 
                 1810 
               
               
                   
                 VH_CK(S176E).001 
                   
                   
                   
               
               
                 377055 
                 104_9C8_CK(S176K)_Fc.001_3.2_VH_CL(S 
                 1812 
                 1814 
                 1816 
               
               
                   
                 176E).001 
                   
                   
                   
               
               
                 377060 
                 105_23B3_VH4_CK(S176K)_Fc.001_Adalimumab_ 
                 1818 
                 1820 
                 1822 
               
               
                   
                 VH_CK(S176E).001 
                   
                   
                   
               
               
                 377064 
                 106_23B3_VH4_CK(S176K)_Fc.001_3.2_V 
                 1824 
                 1826 
                 1828 
               
               
                   
                 H_CL(S176E).001 
                   
                   
                   
               
               
                 377068 
                 117_Adalimumab_IgG.003_3B3v2_VH_CH1 
                 1830 
                 1832 
                 1834 
               
               
                   
                 (S183E).002 
                   
                   
                   
               
               
                 377072 
                 118_Adalimumab_IgG.003_9C8_VH_CH1(S 
                 1836 
                 1838 
                 1840 
               
               
                   
                 183E).002 
                   
                   
                   
               
               
                 377077 
                 119_Adalimumab_IgG.003_23B3_VH4_VH_ 
                 1842 
                 1844 
                 1846 
               
               
                   
                 CH1(S183E).002 
                   
                   
                   
               
               
                 377082 
                 120_3.2_IgG.003_3B3v2_VH_CH1(S183E).002 
                 1848 
                 1850 
                 1852 
               
               
                 377087 
                 121_3.2_IgG.003_9C8_VH_CH1(S183E).002 
                 1854 
                 1856 
                 1858 
               
               
                 377092 
                 122_3.2_IgG.003_23B3_VH4_VH_CH1(S18 
                 1860 
                 1862 
                 1864 
               
               
                   
                 3E).002 
                   
                   
                   
               
               
                 377097 
                 123_3B3v2_IgG.003_Adalimumab_VH_CH1 
                 1866 
                 1868 
                 1870 
               
               
                   
                 (S183E).002 
                   
                   
                   
               
               
                 377102 
                 124_3B3v2_IgG.003_3.2_VH_CH1(S183E).002 
                 1872 
                 1874 
                 1876 
               
               
                 377107 
                 125_9C8_IgG.003_Adalimumab_VH_CH1(S 
                 1878 
                 1880 
                 1882 
               
               
                   
                 183E).002 
                   
                   
                   
               
               
                 3771112 
                 126_9C8_IgG.003_3.2_VH_CH1(S183E).002 
                 1884 
                 1886 
                 1888 
               
               
                 377116 
                 127_23B3_VH4_IgG.003_Adalimumab_VH_ 
                 1890 
                 1892 
                 1894 
               
               
                   
                 CH1(S183E).002 
                   
                   
                   
               
               
                 377121 
                 128_23B3_VH4_IgG.003_3.2_VH_CH1(S18 
                 1896 
                 1898 
                 1900 
               
               
                   
                 3E).002 
                   
                   
                   
               
               
                 377126 
                 129_Adalimumab_CK(S176K)_Fc.002_3B3v 
                 1902 
                 1904 
                 1906 
               
               
                   
                 2_VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377133 
                 130_Adalimumab_CK(S176K)_Fc.002_9C8_ 
                 1908 
                 1910 
                 1912 
               
               
                   
                 VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377138 
                 131_Adalimumab_CK(S176K)_Fc.002_23B3_ 
                 1914 
                 1916 
                 1918 
               
               
                   
                 VH4_VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377142 
                 132_3.2_CL(S176K)_Fc.002_3B3v2_VH_C 
                 1920 
                 1922 
                 1924 
               
               
                   
                 H1(S183E).002 
                   
                   
                   
               
               
                 377148 
                 133_3.2_CL(S176K)_Fc.002_9C8_VH_CH1 
                 1926 
                 1928 
                 1930 
               
               
                   
                 (S183E).002 
                   
                   
                   
               
               
                 377152 
                 134_3.2_CL(S176K)_Fc.002_23B3_VH4_V 
                 1932 
                 1934 
                 1936 
               
               
                   
                 H_CH1(S183E).002 
                   
                   
                   
               
               
                 377156 
                 135_3B3v2_CK(S176K)_Fc.002_Adalimumab_ 
                 1938 
                 1940 
                 1942 
               
               
                   
                 VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377162 
                 136_3B3v2_CK(S176K)_Fc.002_3.2_VH_C 
                 1944 
                 1946 
                 1948 
               
               
                   
                 H1(S183E).002 
                   
                   
                   
               
               
                 377166 
                 137_9C8_CK(S176K)_Fc.002_Adalimumab_ 
                 1950 
                 1952 
                 1954 
               
               
                   
                 VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377172 
                 1389C8_CK(S176K)_Fc.002_3.2_VH_CH1 
                 1956 
                 1958 
                 1960 
               
               
                   
                 (S183E).002 
                   
                   
                   
               
               
                 377176 
                 139_23B3_VH4_CK(S176K)_Fc.002_Adalimumab_ 
                 1962 
                 1964 
                 1966 
               
               
                   
                 VH_CH1(S183E).002 
                   
                   
                   
               
               
                 377182 
                 140_23B3_VH4_CK(S176K)_Fc.002_3.2_V 
                 1968 
                 1970 
                 1972 
               
               
                   
                 H_CH1(S183E).002 
                   
                   
                   
               
               
                 377186 
                 141_Adalimumab_IgG.003_3B3v2_VH_CK 
                 1974 
                 1976 
                 1978 
               
               
                   
                 (S176E).002 
                   
                   
                   
               
               
                 377193 
                 142_Adalimumab_IgG.003_9C8_VH_CK(S1 
                 1980 
                 1982 
                 1984 
               
               
                   
                 76E).002 
                   
                   
                   
               
               
                 377200 
                 143_Adalimumab_IgG.003_23B3_VH4_VH_ 
                 1986 
                 1988 
                 1990 
               
               
                   
                 CK(S176E).002 
                   
                   
                   
               
               
                 377207 
                 144_3.2_IgG.003_3B3v2_VH_CK(S176E).002 
                 1992 
                 1994 
                 1996 
               
               
                 377212 
                 145_3.2_IgG.003_9C8_VH_CK(S176E).002 
                 1998 
                 2000 
                 2002 
               
               
                 377217 
                 146_3.2_IgG.003_23B3_VH4_VH_CK(S176 
                 2004 
                 2006 
                 2008 
               
               
                   
                 E).002 
                   
                   
                   
               
               
                 377222 
                 147_3B3v2_IgG.003_Adalimumab_VH_CK 
                 2010 
                 2012 
                 2014 
               
               
                   
                 (S176E).002 
                   
                   
                   
               
               
                 377229 
                 148_3B3v2_IgG.003_3.2_VH_CL(S176E).002 
                 2016 
                 2018 
                 2020 
               
               
                 377236 
                 149_9C8_IgG.003_Adalimumab_VH_CK(S1 
                 2022 
                 2024 
                 2026 
               
               
                   
                 76E).002 
                   
                   
                   
               
               
                 377241 
                 150_9C8_IgG.003_3.2_VH_CL(S176E).002 
                 2028 
                 2030 
                 2032 
               
               
                 377246 
                 151_23B3_VH4_IgG.003_Adalimumab_VH_ 
                 2034 
                 2036 
                 2038 
               
               
                   
                 CK(S176E).002 
                   
                   
                   
               
               
                 377251 
                 152_23B3_VH4_IgG.003_3.2_VH_CL(S176 
                 2040 
                 2042 
                 2044 
               
               
                   
                 E).002 
                   
                   
                   
               
               
                 377256 
                 153_Adalimumab_CK(S176K)_Fc.002_3B3v 
                 2046 
                 2048 
                 2050 
               
               
                   
                 2_VH_CK(S176E).002 
                   
                   
                   
               
               
                 377261 
                 154_Adalimumab_CK(S176K)_Fc.002_9C8_ 
                 2052 
                 2054 
                 2056 
               
               
                   
                 VH_CK(S176E).002 
                   
                   
                   
               
               
                 377266 
                 155_Adalimumab_CK(S176K)_Fc.002_23B3_ 
                 2058 
                 2060 
                 2062 
               
               
                   
                 VH4_VH_CK(S176E).002 
                   
                   
                   
               
               
                 377271 
                 156_3.2_CL(S176K)_Fc.002_3B3v2_VH_C 
                 2064 
                 2066 
                 2068 
               
               
                   
                 K(S176E).002 
                   
                   
                   
               
               
                 377276 
                 157_3.2_CL(S176K)_Fc.0029C8_VH_CK(S 
                 2070 
                 2072 
                 2074 
               
               
                   
                 176E).002 
                   
                   
                   
               
               
                 377281 
                 158_3.2_CL(S176K)_Fc.002_23B3_VH4_VH_ 
                 2076 
                 2078 
                 2080 
               
               
                   
                 CK(S176E).002 
                   
                   
                   
               
               
                 377286 
                 159_3B3v2_CK(S176K)_Fc.002_Adalimumab_ 
                 2082 
                 2084 
                 2086 
               
               
                   
                 VH_CK(S176E).002 
                   
                   
                   
               
               
                 377290 
                 160_3B3v2_CK(S176K)_Fc.002_3.2_VH_CL 
                 2088 
                 2090 
                 2092 
               
               
                   
                 (S176E).002 
                   
                   
                   
               
               
                 377295 
                 161_9C8_CK(S176K)_Fc.002_Adalimumab_ 
                 2094 
                 2096 
                 2098 
               
               
                   
                 VH_CK(S176E).002 
                   
                   
                   
               
               
                 377300 
                 162_9C8_CK(S176K)_Fc.002_3.2_VH_CL(S 
                 2100 
                 2102 
                 2104 
               
               
                   
                 176E).002 
                   
                   
                   
               
               
                 377305 
                 163_23B3_VH4_CK(S176K)_Fc.002_Adalimumab_ 
                 2106 
                 2108 
                 2110 
               
               
                   
                 VH_CK(S176E).002 
                   
                   
                   
               
               
                 377310 
                 164_23B3_VH4_CK(S176K)_Fc.002_3.2_V 
                 2112 
                 2114 
                 2116 
               
               
                   
                 H_CL(S176E).002 
               
               
                   
               
            
           
         
       
     
     The foregoing amino acid sequences are encoded by the nucleic acid sequences immediately preceding them in the Sequence Listing. 
     The CDR sequences of the VL and VH domains of the foregoing are as shown in Tables 21.2A and 21.2B below. 
     
       
         
           
               
             
               
                 TABLE 21.2A 
               
             
            
               
                   
               
               
                 anti-TL1A/anti-TNFα IgG-Fab 
               
               
                 VL CDR sequences 
               
            
           
           
               
               
               
               
               
            
               
                 Ab 
                 Type 
                 CDRL1 
                 CDRL2 
                 CDRL3 
               
               
                   
               
               
                 Anti-TNF- 
                 NA 
                 CGCGC 
                 GCCGC 
                 CAGAG 
               
               
                 alpha 
                   
                 GTCCC 
                 CTCGA 
                 ATACA 
               
               
                 (Adalimumab) 
                   
                 AGGGA 
                 CTCTT 
                 ACCGA 
               
               
                   
                   
                 ATCCG 
                 CA 
                 GCGC 
               
               
                   
                   
                 GAATT 
                 GAGT 
                 CTTAC 
               
               
                   
                   
                 ACCTC 
                 SEQ ID 
                 ACA 
               
               
                   
                   
                 GCA 
                 NO: 959 
                 SEQ ID 
               
               
                   
                   
                 SEQ ID 
                   
                 NO: 960 
               
               
                   
                   
                 NO: 957 
                   
                   
               
               
                   
               
               
                   
                 AA 
                 RASQG 
                 AASTL 
                 QRYNR 
               
               
                   
                   
                 IRNYL 
                 QS 
                 APYT 
               
               
                   
                   
                 A 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                   
                 SEQ ID 
                 NO: 242 
                 NO: 96 
               
               
                   
                   
                 NO: 92 
                   
                   
               
               
                   
               
               
                 Anti-TNF- 
                 NA 
                 ACTGG 
                 GGTAA 
                 CAGTC 
               
               
                 alpha (3.2) 
                   
                 GAGCA 
                 CAGCA 
                 CTATG 
               
               
                   
                   
                 GTTCC 
                 ATCGG 
                 ACAGC 
               
               
                   
                   
                 AACAT 
                 CCCTC 
                 AGCC 
               
               
                   
                   
                 CG 
                 A 
                 TGAGT 
               
               
                   
                   
                 GGGCA 
                 SEQ ID 
                 GGTTC 
               
               
                   
                   
                 GGTTA 
                 NO: 965 
                 GGTG 
               
               
                   
                   
                 TGATG 
                   
                 SEQ ID 
               
               
                   
                   
                 TACAC 
                   
                 NO: 
               
               
                   
                   
                 SEQ ID 
                   
                 1189 
               
               
                   
                   
                 NO: 962 
                   
                   
               
               
                   
               
               
                   
                 AA 
                 TGSSS 
                 GNSNR 
                 QSYDS 
               
               
                   
                   
                 NIGAG 
                 PS 
                 SLSGS 
               
               
                   
                   
                 YDVH 
                 SEQ ID 
                 V 
               
               
                   
                   
                 SEQ ID 
                 NO: 148 
                 SEQ ID 
               
               
                   
                   
                 NO: 146 
                   
                 NO: 150 
               
               
                   
               
               
                 Anti-TL1A 
                 NA 
                 AGGGC 
                 GGTGC 
                 CAGCA 
               
               
                 (3B3v2) 
                   
                 CAGTC 
                 ATCCA 
                 GTATG 
               
               
                   
                   
                 AGAGT 
                 GCAGG 
                 GTAGC 
               
               
                   
                   
                 GTTAG 
                 GC 
                 TCAC 
               
               
                   
                   
                 AA 
                 CACT 
                 CTACC 
               
               
                   
                   
                 GCAGT 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                   
                 TACTT 
                 NO: 467 
                 NO: 104 
               
               
                   
                   
                 AGCC 
                   
                   
               
               
                   
                   
                 SEQ ID 
                   
                   
               
               
                   
                   
                 NO: 94 
                   
                   
               
               
                   
               
               
                   
                 AA 
                 RASQS 
                 GASSR 
                 QQYGS 
               
               
                   
                   
                 VRSSY 
                 AT 
                 SFT 
               
               
                   
                   
                 LA 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                   
                 SEQ ID 
                 NO: 124 
                 NO: 126 
               
               
                   
                   
                 NO: 122 
                   
                   
               
               
                   
               
               
                 Anti-TL1A 
                 NA 
                 CGGGC 
                 GCTGC 
                 CAACA 
               
               
                 (9C8) 
                   
                 AAGTC 
                 ATCCA 
                 GAGTT 
               
               
                   
                   
                 AGAGC 
                 GTTTG 
                 ACAGT 
               
               
                   
                   
                 ATTAA 
                 CA 
                 ACCC 
               
               
                   
                   
                 CA 
                 AAGT 
                 CTCGG 
               
               
                   
                   
                 ACTAT 
                 SEQ ID 
                 ACG 
               
               
                   
                   
                 TTAAA 
                 NO: 2117 
                 SEQ ID 
               
               
                   
                   
                 T 
                   
                 NO: 2118 
               
               
                   
                   
                 SEQ ID 
                   
                   
               
               
                   
                   
                 NO: 263 
                   
                   
               
               
                   
               
               
                   
                 AA 
                 RASQS 
                 AASSL 
                 QQSYS 
               
               
                   
                   
                 INNYL 
                 QS 
                 TPRT 
               
               
                   
                   
                 N 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                   
                 SEQ ID 
                 NO: 112 
                 NO: 108 
               
               
                   
                   
                 NO: 110 
                   
                   
               
               
                   
               
               
                 Anti-TL1A 
                 NA 
                 AGGTC 
                 TGGGC 
                 CAGCA 
               
               
                 (23B3) 
                   
                 CAGCC 
                 ATCTA 
                 ATATT 
               
               
                   
                   
                 AGAGT 
                 CCCGG 
                 ATAAG 
               
               
                   
                   
                 GTGTT 
                 GA 
                 ACTC 
               
               
                   
                   
                 AT 
                 ATCC 
                 CTCTC 
               
               
                   
                   
                 ACAGC 
                 SEQ ID 
                 ACT 
               
               
                   
                   
                 TCCAA 
                 NO: 
                 SEQ ID 
               
               
                   
                   
                 CAATA 
                 2120 
                 NO: 2121 
               
               
                   
                   
                 AGAAC 
                   
                   
               
               
                   
                   
                 TA 
                   
                   
               
               
                   
                   
                 CTTAG 
                   
                   
               
               
                   
                   
                 TT 
                   
                   
               
               
                   
                   
                 SEQ ID 
                   
                   
               
               
                   
                   
                 NO: 
                   
                   
               
               
                   
                   
                 2119 
                   
                   
               
               
                   
               
               
                   
                 AA 
                 RSSQS 
                 WASTR 
                 QQYYK 
               
               
                   
                   
                 VLYSS 
                 ES 
                 TPLT 
               
               
                   
                   
                 NNKNY 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                   
                 LV 
                 NO: 118 
                 NO: 120 
               
               
                   
                   
                 SEQ ID 
                   
                   
               
               
                   
                   
                 NO: 128 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 21.2B: 
               
             
            
               
                   
               
               
                 anti-TL1A/anti-TNF-α IgG-Fab 
               
               
                 VH CDR sequences 
               
            
           
           
               
               
               
               
               
            
               
                 Ab 
                 Type 
                 CDRH1 
                 CDRH2 
                 CDRH3 
               
               
                   
               
               
                 Anti-TNF- 
                 NA 
                 GATTA 
                 GCCAT 
                 GTCTC 
               
               
                 alpha 
                   
                 TGCGA 
                 TACGT 
                 CTACT 
               
               
                 (Adalimumab) 
                   
                 TGCAT 
                 GGAAT 
                 TGTCT 
               
               
                   
                   
                 SEQ ID 
                 AGCGG 
                 ACAGC 
               
               
                   
                   
                 NO: 2122 
                 ACACA 
                 TTCGT 
               
               
                   
                   
                   
                 TCGAT 
                 CGCTC 
               
               
                   
                   
                   
                 TATGC 
                 GACTA 
               
               
                   
                   
                   
                 AGACA 
                 T 
               
               
                   
                   
                   
                 GTGTG 
                 SEQ ID 
               
               
                   
                   
                   
                 GAGGG 
                 NO: 2124 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 2123 
                   
               
               
                   
               
               
                   
                 AA 
                 DYAMH 
                 AITWN 
                 VSYLS 
               
               
                   
                   
                 SEQ ID 
                 SGHID 
                 TASSL 
               
               
                   
                   
                 NO: 158 
                 YADSV 
                 DY 
               
               
                   
                   
                   
                 EG 
                 SEQ ID 
               
               
                   
                   
                   
                 SEQ ID 
                 NO: 162 
               
               
                   
                   
                   
                 NO: 160 
                   
               
               
                   
               
               
                 Anti-TNF- 
                 NA 
                 AGCTA 
                 ATCAT 
                 AGTAA 
               
               
                 alpha (3.2) 
                   
                 CTGGA 
                 CTATC 
                 CTGGG 
               
               
                   
                   
                 TCGGC 
                 TTGGT 
                 GTCTT 
               
               
                   
                   
                 SEQ ID 
                 GACTC 
                 GACTA 
               
               
                   
                   
                 NO: 2125 
                 AGATA 
                 C 
               
               
                   
                   
                   
                 CCAGA 
                 SEQ ID 
               
               
                   
                   
                   
                 TACAG 
                 NO: 2127 
               
               
                   
                   
                   
                 CCCGT 
                   
               
               
                   
                   
                   
                 CCTTC 
                   
               
               
                   
                   
                   
                 CAAGG 
                   
               
               
                   
                   
                   
                 C 
                   
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 2126 
                   
               
               
                   
               
               
                   
                 AA 
                 SYWIG 
                 IIYLG 
                 SNWGL 
               
               
                   
                   
                 SEQ ID 
                 DSDTR 
                 DY 
               
               
                   
                   
                 NO: 212 
                 YSPSF 
                 SEQ ID 
               
               
                   
                   
                   
                 QG 
                 NO: 216 
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 214 
                   
               
               
                   
               
               
                 Anti-T1A 
                 NA 
                 GGTTA 
                 GAAAT 
                 GGATA 
               
               
                 (3B3v2) 
                   
                 CTACT 
                 CAATC 
                 TTGTA 
               
               
                   
                   
                 GGAAC 
                 ATGCT 
                 GAAGT 
               
               
                   
                   
                 SEQ ID 
                 GGAAA 
                 ACCAC 
               
               
                   
                   
                 NO: 2128 
                 CACCA 
                 CTGCT 
               
               
                   
                   
                   
                 ACTAC 
                 ACTTT 
               
               
                   
                   
                   
                 AACCC 
                 GACTA 
               
               
                   
                   
                   
                 GTCCC 
                 C 
               
               
                   
                   
                   
                 TCAAG 
                 SEQ ID 
               
               
                   
                   
                   
                 AGT 
                 NO: 2130 
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 2129 
                   
               
               
                   
               
               
                   
                 AA 
                 GYYWN 
                 EINHA 
                 GYCRS 
               
               
                   
                   
                 SEQ ID 
                 GNTNY 
                 TTCYF 
               
               
                   
                   
                 NO: 188 
                 NPSLK 
                 DY 
               
               
                   
                   
                   
                 S 
                 SEQ ID 
               
               
                   
                   
                   
                 SEQ ID 
                 NO: 192 
               
               
                   
                   
                   
                 NO: 190 
                   
               
               
                   
               
               
                 Anti-TL1A 
                 NA 
                 AGTTA 
                 TATAT 
                 GAAAC 
               
               
                 (9C8) 
                   
                 CTTCT 
                 CTATT 
                 TGGGA 
               
               
                   
                   
                 GGAGC 
                 ACAGT 
                 GCTAC 
               
               
                   
                   
                 SEQ ID 
                 GGGCA 
                 TACGG 
               
               
                   
                   
                 NO: 2131 
                 GACCA 
                 CTTTG 
               
               
                   
                   
                   
                 AATAC 
                 ACTAC 
               
               
                   
                   
                   
                 AACCC 
                 SEQ ID 
               
               
                   
                   
                   
                 CTCCC 
                 NO: 2133 
               
               
                   
                   
                   
                 TCAAG 
                   
               
               
                   
                   
                   
                 AGT 
                   
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 2132 
                   
               
               
                   
               
               
                   
                 AA 
                 SYFWS 
                 YIYYS 
                 ETGSY 
               
               
                   
                   
                 SEQ ID 
                 GQTKY 
                 YGFDY 
               
               
                   
                   
                 NO: 170 
                 NPSLK 
                 SEQ ID 
               
               
                   
                   
                   
                 S 
                 NO: 180 
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 633 
                   
               
               
                   
               
               
                 Anti-TL1A 
                 NA 
                 ACCAA 
                 AGGAC 
                 GAGGA 
               
               
                 (23B3) 
                   
                 CAGTG 
                 ATACT 
                 TGGGG 
               
               
                   
                   
                 TTGCT 
                 ACAGG 
                 ATAGC 
               
               
                   
                   
                 TGGAA 
                 TCCAA 
                 TACTA 
               
               
                   
                   
                 C 
                 GTGGT 
                 CCGCT 
               
               
                   
                   
                 SEQ ID 
                 ATAAT 
                 ACGGT 
               
               
                   
                   
                 NO: 2134 
                 GATTA 
                 ATGGA 
               
               
                   
                   
                   
                 TGCAG 
                 CGTC 
               
               
                   
                   
                   
                 TTTCT 
                 SEQ ID 
               
               
                   
                   
                   
                 CTGAA 
                 NO: 2136 
               
               
                   
                   
                   
                 AAGT 
                   
               
               
                   
                   
                   
                 SEQ ID 
                   
               
               
                   
                   
                   
                 NO: 2135 
                   
               
               
                   
               
               
                   
                 AA 
                 TNSVA 
                 RTYYR 
                 EDGDS 
               
               
                   
                   
                 WN 
                 SKWYN 
                 YYRYG 
               
               
                   
                   
                 SEQ ID 
                 DYAVS 
                 MDV 
               
               
                   
                   
                 NO: 182 
                 LKS 
                 SEQ ID 
               
               
                   
                   
                   
                 SEQ ID 
                 NO: 186 
               
               
                   
                   
                   
                 NO: 196 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 21.3 
               
             
            
               
                   
               
               
                 Titer, Recovery, and Activity of IgG-Fabs 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                   
                 hu  
                 hu TL1A 
                 hu  
                 hu TNFa 
               
               
                 iPS  
                   
                   
                 CL/CH1 
                   
                 Titer 
                 Recovery 
                 TL1A 
                 Fold 
                 TNFa 
                 Fold 
               
               
                 No. 
                 IgG 
                 Fab 
                 Swap 
                 CPM 
                 (mg/L) 
                 (mg/L) 
                 (pM) 
                 Change 
                 (pM) 
                 Change 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                 376597 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v1 
                 63.5 
                 56.26698 
                 197.3 
                 3.653704 
                 87.5 
                 0.603448 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376601 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v1 
                 49.4 
                 44.46469 
                 136.5 
                 0.903974 
                 96.5 
                 0.665517 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376605 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v1 
                 46 
                 37.35632 
                 689.6 
                 5.56129 
                 941.7 
                 6.494483 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376609 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v1 
                 28.9 
                 37.88288 
                 4408 
                 81.62963 
                 94.2 
                 1.345714 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376613 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v1 
                 34.5 
                 41.14599 
                 260.3 
                 1.723841 
                 75.3 
                 1.075714 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376617 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v1 
                 30.3 
                 37.13529 
                 1025 
                 8.266129 
                 79.6 
                 1.137143 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376621 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v1 
                 53.1 
                 55.23171 
                 66.3 
                 1.227778 
                 33.3 
                 0.229655 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376625 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v1 
                 51.2 
                 62.13592 
                 63.8 
                 1.181481 
                 37.3 
                 0.532857 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376629 
                 TL1A (9C8) 
                 TNFa (ada) 
                 No swap 
                 v1 
                 46.1 
                 43.10706 
                 142.1 
                 0.94106 
                 22.4 
                 0.154483 
               
               
                 376633 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 No swap 
                 v1 
                 44.3 
                 53.37693 
                 215.7 
                 1.428477 
                 44.6 
                 0.637143 
               
               
                 376637 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v1 
                 45.9 
                 43.74483 
                 348.8 
                 2.812903 
                 27.5 
                 0.189655 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376641 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v1 
                 49.7 
                 56.93 
                 208.8 
                 1.683871 
                 43.9 
                 0.627143 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376645 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v1 
                 5.82 
                 4.959868 
                 139.1 
                 2.575926 
                 131 
                 0.903448 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376651 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v1 
                 5.73 
                 7.33602 
                 337.8 
                 2.237086 
                 157.5 
                 1.086207 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376655 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v1 
                 6.54 
                 6.945786 
                 200.2 
                 1.614516 
                 198.5 
                 1.368966 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376659 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v1 
                 7.33 
                 7.576692 
                 73.6 
                 1.362963 
                 92.4 
                 1.32 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376665 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v1 
                 18.3 
                 20.86174 
                 91 
                 0.602649 
                 41.7 
                 0.595714 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376669 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v1 
                 21.8 
                 21.92793 
                 77.6 
                 0.625806 
                 73.5 
                 1.05 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376673 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v1 
                 27.3 
                 27.46576 
                 4825 
                 89.35185 
                 41.7 
                 0.287586 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376679 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v1 
                 28.8 
                 40.93307 
                 6250 
                 115.7407 
                 39.6 
                 0.565714 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376683 
                 TL1A (9C8) 
                 TNFa (ada) 
                 N-term 
                 v1 
                 33.4 
                 28.83716 
                 414 
                 2.741722 
                 35.2 
                 0.242759 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376689 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 N-term 
                 v1 
                 32.4 
                 39.77615 
                 787.1 
                 5.212583 
                 45 
                 0.642857 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376693 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v1 
                 25.4 
                 19.95322 
                   
                   
                 29.5 
                 0.203448 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376699 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v1 
                 22.9 
                 25.85043 
                   
                   
                 40.7 
                 0.581429 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376703 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v1 
                 39.2 
                 30.58153 
                 150.4 
                 2.785185 
                 78.6 
                 0.542069 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376709 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v1 
                 50 
                 41.99633 
                 199.5 
                 1.321192 
                 80.4 
                 0.554483 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376715 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v1 
                 40.7 
                 29.97569 
                 640.4 
                 5.164516 
                 64.2 
                 0.442759 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376721 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v1 
                 23.2 
                 24.91097 
                 213.2 
                 3.948148 
                 92.8 
                 1.325714 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376725 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v1 
                 26.2 
                 31.24415 
                 185.8 
                 1.230464 
                 48.3 
                 0.69 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376729 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v1 
                 31.7 
                 28.69498 
                 330.8 
                 2.667742 
                 79 
                 1.128571 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376733 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v1 
                 36.9 
                 31.95548 
                 66.1 
                 1.224074 
                 263 
                 1.813793 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376739 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v1 
                 2.95 
                 1.921946 
                   
                   
                   
                   
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376745 
                 TL1A (9C8) 
                 TNFa (ada) 
                 C-term 
                 v1 
                 24.9 
                 22.1839 
                 108.7 
                 0.719868 
                 135.3 
                 0.933103 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376750 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 C-term 
                 v1 
                 7.71 
                 8.223697 
                 223 
                 1.476821 
                 71.4 
                 1.02 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376755 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v1 
                 9.11 
                 7.361222 
                 271.5 
                 2.189516 
                 433.3 
                 2.988276 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376760 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v1 
                 7.17 
                 5.960717 
                 305.6 
                 2.464516 
                 129.8 
                 1.854286 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376765 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v1 
                 3.66 
                 4.304958 
                 343.6 
                 6.362963 
                 182 
                 1.255172 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376770 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v1 
                 2.68 
                 1.712581 
                   
                   
                   
                 0 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376775 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v1 
                 4.99 
                 2.967115 
                 389.7 
                 3.142742 
                 90.4 
                 0.623448 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376779 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v1 
                 4.92 
                 3.717088 
                 707.1 
                 13.09444 
                 226.8 
                 3.24 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376785 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v1 
                 9.36 
                 9.546385 
                 149.5 
                 0.990066 
                 42 
                 0.6 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376789 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v1 
                 12.2 
                 11.32345 
                 554.2 
                 4.469355 
                 74 
                 1.057143 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376793 
                 (3B3v2) 
                 TL1A 
                 Both  
                 v1 
                 19.3 
                 15.35021 
                 1937.5 
                 35.87963 
                 347.9 
                 2.39931 
               
               
                   
                   
                 TNFa (ada) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376797 
                 (3B3v2) 
                 TL1A 
                 Both  
                 v1 
                 1 
                 0 
                   
                   
                   
                   
               
               
                   
                   
                 TNFa (3.2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376801 
                 TL1A (9C8) 
                 TNFa (ada) 
                 Both  
                 v1 
                 13.6 
                 11.14213 
                 286.8 
                 1.899338 
                 310 
                 2.137931 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376806 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 Both  
                 v1 
                 1 
                 0.961822 
                   
                   
                   
                   
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376811 
                 TL1A 
                 TNFa (ada) 
                 Both  
                 v1 
                 3.22 
                 1.671524 
                   
                   
                   
                   
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376817 
                 TL1A 
                 TNFa (3.2) 
                 Both  
                 v1 
                 1 
                 0 
                   
                   
                   
                   
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376822 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v2 
                 53.6 
                 43.05822 
                 128.5 
                 2.37963 
                 84.7 
                 0.584138 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376826 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v2 
                 27.4 
                 22.92398 
                 106 
                 0.701987 
                 80.8 
                 0.557241 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376830 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v2 
                 34.2 
                 25.0415 
                 262 
                 2.112903 
                 73.3 
                 0.505517 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376834 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v2 
                 18 
                 17.62724 
                 246.8 
                 4.57037 
                 97 
                 1.385714 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376838 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v2 
                 17.3 
                 17.06775 
                 548.8 
                 3.634437 
                 31.6 
                 0.451429 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376842 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v2 
                 14.9 
                 12.78592 
                 469.85 
                 3.789113 
                 82.9 
                 1.184286 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376846 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v2 
                 46.7 
                 45.57382 
                 71.8 
                 1.32963 
                 31.5 
                 0.217241 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376850 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v2 
                 24.4 
                 25.47009 
                 83.3 
                 1.542593 
                 84.8 
                 1.211429 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376854 
                 TL1A (9C8) 
                 TNFa (ada) 
                 No swap 
                 v2 
                 39.5 
                 36.01855 
                 229.3 
                 1.518543 
                 30.8 
                 0.212414 
               
               
                 376858 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 No swap 
                 v2 
                 17.5 
                 17.37068 
                 190.8 
                 1.263576 
                 35 
                 0.5 
               
               
                 376862 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v2 
                 23.5 
                 19.86708 
                 436.3 
                 3.518548 
                 34.4 
                 0.237241 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376867 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v2 
                 11.3 
                 10.11778 
                 109 
                 0.879032 
                 29 
                 0.414286 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376872 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v2 
                 5.26 
                 5.362855 
                 27.725 
                 0.513426 
                 519.5 
                 3.582759 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376879 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v2 
                 4.24 
                 2.812933 
                 253.2 
                 1.676821 
                 173.2 
                 1.194483 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376885 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v2 
                 3.01 
                 2.592265 
                 310.3 
                 2.502419 
                 940.8 
                 6.488276 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376889 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v2 
                 1 
                 0.817101 
                   
                   
                   
                   
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376897 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v2 
                 2.76 
                 5.494626 
                 760 
                 5.033113 
                 472.9 
                 6.755714 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376902 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v2 
                 1 
                 0.985458 
                   
                   
                   
                   
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376907 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v2 
                 36.7 
                 29.31227 
                 3213 
                 59.5 
                 35.8 
                 0.246897 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376913 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v2 
                 19.4 
                 25.22426 
                 813 
                 15.05556 
                 52.55 
                 0.750714 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376918 
                 TL1A (9C8) 
                 TNFa (ada) 
                 N-term 
                 v2 
                 39.6 
                 31.18441 
                 203.6 
                 1.348344 
                 17.855 
                 0.123138 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376925 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 N-term 
                 v2 
                 22.4 
                 27.25854 
                 189.65 
                 1.25596 
                 29.35 
                 0.419286 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376931 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v2 
                 22.9 
                 20.43729 
                 7195 
                 58.02419 
                 26.77 
                 0.184621 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376937 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v2 
                 12.2 
                 9.722544 
                   
                   
                 176.95 
                 2.527857 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376941 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v2 
                 40.6 
                 31.79866 
                 22.73 
                 0.420926 
                 50.4 
                 0.347586 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376949 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v2 
                 38.7 
                 35.043 
                 48.165 
                 0.318974 
                 49.095 
                 0.338586 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376955 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v2 
                 30 
                 24.1244 
                 333.95 
                 2.693145 
                 57.15 
                 0.394138 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376962 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v2 
                 9.96 
                 13.79268 
                 540 
                 10 
                 1957 
                 27.95714 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376967 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v2 
                 8.73 
                 12.01796 
                   
                   
                   
                   
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376972 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v2 
                 2.58 
                 4.419175 
                 12490 
                 100.7258 
                 7870 
                 112.4286 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376976 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v2 
                 26.3 
                 16.32049 
                 31.31 
                 0.579815 
                 172.2 
                 1.187586 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376983 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v2 
                 1 
                 0.993437 
                   
                   
                   
                   
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376991 
                 TL1A (9C8) 
                 TNFa (ada) 
                 C-term 
                 v2 
                 40.5 
                 38.42849 
                 59.9 
                 0.396689 
                 148 
                 1.02069 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 376995 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 C-term 
                 v2 
                 1 
                 0.339488 
                   
                   
                   
                   
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377001 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v2 
                 25.8 
                 25.49058 
                 41.675 
                 0.336089 
                 158.45 
                 1.092759 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377006 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v2 
                 1 
                 0.124147 
                   
                   
                   
                   
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377011 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v2 
                 1 
                 3.586728 
                 2702 
                 50.03704 
                 3065.5 
                 21.14138 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377015 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v2 
                 3.2 
                 3.312442 
                   
                   
                 141050 
                 972.7586 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377020 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v2 
                 1 
                 1.15137 
                   
                   
                   
                   
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377025 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v2 
                 1 
                 0.778064 
                   
                   
                   
                   
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377031 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v2 
                 1 
                 2.940239 
                 17640 
                 116.8212 
                 12965 
                 185.2143 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377036 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v2 
                 1 
                 0.595615 
                   
                   
                   
                   
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377041 
                 TL1A 
                 TNFa (ada) 
                 Both  
                 v2 
                 10.8 
                 10.82989 
                 1689.5 
                 31.28704 
                 2014.5 
                 13.8931 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377045 
                 TL1A 
                 TNFa (3.2) 
                 Both  
                 v2 
                 1 
                 0 
                   
                   
                   
                   
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377050 
                 TL1A (9C8) 
                 TNFa (ada) 
                 Both  
                 v2 
                 22.1 
                 16.88003 
                 269.05 
                 1.781788 
                 67.2 
                 0.463448 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377056 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 Both  
                 v2 
                 1 
                 0 
                   
                   
                   
                   
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377060 
                 TL1A 
                 TNFa (ada) 
                 Both  
                 v2 
                 1 
                 3.78306 
                 2371 
                 19.12097 
                 171.5 
                 1.182759 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377064 
                 TL1A 
                 TNFa (3.2) 
                 Both  
                 v2 
                 1 
                 0 
                   
                   
                   
                   
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377068 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v3 
                 69.2 
                 61.35414 
                 23.5 
                 0.435185 
                 56.1 
                 0.386897 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377073 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v3 
                 64.9 
                 57.53099 
                 47.08 
                 0.311788 
                 36.795 
                 0.253759 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377077 
                 TNFa (ada) 
                 TL1A 
                 No swap 
                 v3 
                 60 
                 50.44568 
                 67.9 
                 0.547581 
                 49.66 
                 0.342483 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377082 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v3 
                 35.6 
                 50.01275 
                 194.9 
                 3.609259 
                 1213.5 
                 17.33571 
               
               
                   
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377087 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v3 
                 44.7 
                 33 18984 
                 148.8 
                 0.98543 
                 41.555 
                 0.593643 
               
               
                   
                   
                 (9C8) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377092 
                 TNFa (3.2) 
                 TL1A 
                 No swap 
                 v3 
                 35 
                 43.58861 
                 598.5 
                 4.826613 
                 38.845 
                 0.554929 
               
               
                   
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377097 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v3 
                 70.5 
                 74.25303 
                 37.29 
                 0.690556 
                 20.85 
                 0.143793 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377102 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v3 
                 68.1 
                 84.23872 
                 31.565 
                 0.584537 
                 29.98 
                 0.428286 
               
               
                   
                 (3B3v2) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377107 
                 TL1A (9C8) 
                 TNFa (ada) 
                 No swap 
                 v3 
                 57 
                 55.22568 
                 78.35 
                 0.518874 
                 16.775 
                 0.11569 
               
               
                 377112 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 No swap 
                 v3 
                 37.9 
                 45.76692 
                 53.5 
                 0.354305 
                 26.315 
                 0.375929 
               
               
                 377116 
                 TL1A 
                 TNFa (ada) 
                 No swap 
                 v3 
                 44.3 
                 44.02405 
                 73.9 
                 0.595968 
                 15.23 
                 0.105034 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377121 
                 TL1A 
                 TNFa (3.2) 
                 No swap 
                 v3 
                 40.7 
                 28.59812 
                 62.55 
                 0.504435 
                 22.85 
                 0.326429 
               
               
                   
                 (23B3) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377126 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v3 
                 1 
                 1.981845 
                   
                   
                   
                   
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377133 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v3 
                 2.65 
                 2.948434 
                 806.5 
                 5.34106 
                 552 
                 3.806897 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377138 
                 TNFa (ada) 
                 TL1A 
                 N-term 
                 v3 
                 1 
                 1.33942 
                   
                   
                   
                   
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377142 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v3 
                 10.2 
                 11.75934 
                 32.78 
                 0.607037 
                 45.34 
                 0.647714 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377148 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v3 
                 11.1 
                 13.07635 
                 99.65 
                 0.659934 
                 48.495 
                 0.692786 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377152 
                 TNFa (3.2) 
                 TL1A 
                 N-term 
                 v3 
                 13 
                 12.69725 
                 46.51 
                 0.375081 
                 39.74 
                 0.567714 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377156 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v3 
                 30.8 
                 34.83478 
                 1584.5 
                 29.34259 
                 19.9 
                 0.137241 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377162 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v3 
                 26.2 
                 34.40556 
                 2504 
                 46.37037 
                 24.875 
                 0.355357 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377166 
                 TL1A (9C8) 
                 TNFa (ada) 
                 N-term 
                 v3 
                 31.9 
                 26.76967 
                 83 
                 0.549669 
                 16.955 
                 0.116931 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377172 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 N-term 
                 v3 
                 31.3 
                 38.70441 
                 81.5 
                 0.539735 
                 27.26 
                 0.389429 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377176 
                 TL1A 
                 TNFa (ada) 
                 N-term 
                 v3 
                 11.3 
                 11.48623 
                 5305 
                 42.78226 
                 28.73 
                 0.198138 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377182 
                 TL1A 
                 TNFa (3.2) 
                 N-term 
                 v3 
                 14 
                 15.5418 
                 28185 
                 227.2984 
                 48.17 
                 0.688143 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377186 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v3 
                 46.4 
                 36.6753 
                 37.57 
                 0.695741 
                 35.75 
                 0.246552 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377193 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v3 
                 32 
                 28.52527 
                 128.05 
                 0.848013 
                 30.93 
                 0.21331 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377200 
                 TNFa (ada) 
                 TL1A 
                 C-term 
                 v3 
                 56.6 
                 43.6029 
                 217.35 
                 1.752823 
                 38.425 
                 0.265 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377208 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v3 
                 22.1 
                 26 10432 
                 212.35 
                 3.932407 
                 170.15 
                 2.430714 
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377212 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v3 
                 17.6 
                 18.67336 
                 874 
                 5.788079 
                 306.8 
                 4.382857 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377217 
                 TNFa (3.2) 
                 TL1A 
                 C-term 
                 v3 
                 27.4 
                 25.63219 
                 271.4 
                 2.18871 
                 114.8 
                 1.64 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377223 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v3 
                 40.5 
                 36.07182 
                 30.175 
                 0.558796 
                 167 
                 1.151724 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377229 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v3 
                 3.05 
                 3.031093 
                 79.5 
                 1.472222 
                 1711.5 
                 24.45 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377236 
                 TL1A (9C8) 
                 TNFa (ada) 
                 C-term 
                 v3 
                 48.3 
                 43 315 
                 54.25 
                 0.359272 
                 61.7 
                 0.425517 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377241 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 C-term 
                 v3 
                 8.95 
                 8.490845 
                 606 
                 4.013245 
                 268.4 
                 3.834286 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377246 
                 TL1A 
                 TNFa (ada) 
                 C-term 
                 v3 
                 22.4 
                 20.52445 
                 68.7 
                 0.554032 
                 118.5 
                 0.817241 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377252 
                 TL1A 
                 TNFa (3.2) 
                 C-term 
                 v3 
                 5.05 
                 5.690246 
                 219.8 
                 1.772581 
                 1381 
                 19.72857 
               
               
                   
                 (23B3) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377256 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v3 
                 1 
                 3.203042 
                   
                   
                   
                   
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377261 
                 TNFa (ada) 
                 TL1A 
                 Both  
                 v3 
                 1 
                 3.032322 
                   
                   
                   
                   
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377266 
                 TNFa (ada) 
                 TL1A 
                 Both 
                 v3 
                 2.57 
                 3.395799 
                   
                   
                   
                   
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377272 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v3 
                 1 
                 3.071647 
                 13300 
                 246.2963 
                   
                   
               
               
                   
                   
                 (3B3v2) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377276 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v3 
                 14 
                 16.05846 
                 1237.5 
                 8.195364 
                 362.4 
                 5.177143 
               
               
                   
                   
                 (9C8) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377281 
                 TNFa (3.2) 
                 TL1A 
                 Both  
                 v3 
                 12.6 
                 13.70576 
                 1937.5 
                 15.625 
                 282.35 
                 4.033571 
               
               
                   
                   
                 (23B3) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377286 
                 TL1A 
                 TNFa (ada) 
                 Both  
                 v3 
                 25.5 
                 29.40929 
                 486.7 
                 9.012963 
                 521.5 
                 3.596552 
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377290 
                 TL1A 
                 TNFa (3.2) 
                 Both  
                 v3 
                 1 
                 0.304074 
                   
                   
                   
                   
               
               
                   
                 (3B3v2) 
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377295 
                 TL1A (9C8) 
                 TNFa (ada) 
                 Both  
                 v3 
                 26.3 
                 29.74724 
                 177.55 
                 1.175828 
                 132.35 
                 0.912759 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377300 
                 TL1A (9C8) 
                 TNFa (3.2) 
                 Both  
                 v3 
                 3.58 
                 4.355968 
                 17575 
                 116.3907 
                 69750 
                 996.4286 
               
               
                   
                   
                   
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377305 
                 (23B3) 
                 TL1A 
                 Both  
                 v3 
                 11 
                 13.60209 
                   
                   
                 595.5 
                 4.106897 
               
               
                   
                   
                 TNFa (ada) 
                 swap 
                   
                   
                   
                   
                   
                   
                   
               
               
                 377310 
                 (23B3) 
                 TL1A 
                 Both  
                 v3 
                 1 
                 0.64267 
                   
                   
                   
                   
               
               
                   
                   
                 TNFa (3.2) 
                 swap 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 21.4 
               
             
            
               
                   
               
               
                 anti-TL1A/anti-TNF-α IgG-Fab purity 
               
            
           
           
               
               
               
            
               
                   
                   
                 Caliper 
               
            
           
           
               
               
               
               
            
               
                   
                 SE-HPLC 
                 Non-reduced 
                 Reduced 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 iPS No. 
                 pre-MP HMW 
                 MP 
                 post-MP 
                 MP 1 
                 MP 2 
                 MP 3 
                 LMW 
                 HC 1 
                 HC 2 
                 HC  
                 LC 1 
                 R LC 2 
               
               
                   
               
               
                 376597 
                  3.832 
                 96 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.206897 
                 0.793103 
               
               
                 376601 
                  2.894 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.35316  
                 0.64684  
               
               
                 376605 
                  3.362 
                 96.634 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376609 
                  1.241 
                 99 
                   
                 1     
                   
                   
                 0     
                 0.153846 
                 0.846154 
                   
                 1     
                   
               
               
                 376613 
                  1.838 
                 98 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.298658 
                 0.701342 
               
               
                 376617 
                  1.162 
                 99 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376621 
                  1.827 
                 98.172 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0     
                 1     
               
               
                 376625 
                  2.552 
                 97 
                   
                 1     
                   
                   
                 0     
                 0.136503 
                 0.863497 
                   
                 0.511494 
                 0.488506 
               
               
                 376629 
                  1.868 
                 98 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.521429 
                 0.478571 
               
               
                 376633 
                  5.854 
                 94 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.454225 
                 0.545775 
               
               
                 376637 
                  1.099 
                 98.901 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.683333 
                 0.316667 
               
               
                 376641 
                  3.255 
                 96.745 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.44898  
                 0.55102  
               
               
                 376645 
                  8.844 
                 63.754 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.336066 
                 0.663934 
               
               
                 376651 
                 39.874 
                 60 
                   
                 0.46   
                 0.54   
                   
                 0     
                 1     
                   
                   
                 0.364929 
                 0.635071 
               
               
                 376655 
                  8.191 
                 91.809 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.335052 
                 0.664948 
               
               
                 376659 
                  3.515 
                 96.485 
                   
                 0.91   
                   
                   
                 0.09   
                 0.07438 
                 0.92562  
                   
                 0.514599 
                 0.485401 
               
               
                 376665 
                  2.981 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.498168 
                 0.501832 
               
               
                 376669 
                  2.213 
                 97.787 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376673 
                  4.635 
                 60.842 
                   
                 0.95   
                   
                   
                 0.05   
                 0.110115 
                 0.889885 
                   
                 1     
                   
               
               
                 376679 
                  1.774 
                 98.226 
                   
                 0.7    
                   
                   
                 0.3    
                 0.123494 
                 0.203008 
                 0.673499 
                 1     
                   
               
               
                 376683 
                  5.787 
                 94 
                   
                 0.53   
                 0.22   
                   
                 0.25   
                 1     
                   
                   
                 0.330189 
                 0.669811 
               
               
                 376689 
                  3.896 
                 96.104 
                   
                 0.7    
                   
                   
                 0.3    
                 1     
                   
                   
                 0.209386 
                 0.790614 
               
               
                 376693 
                 2.32 
                 96.131 
                  1.548 
                 0.8    
                   
                   
                 0.1    
                 1     
                   
                   
                 1     
                   
               
               
                 376699 
                 1.27 
                 99 
                   
                 0.715   
                   
                   
                 0.285   
                 1     
                   
                   
                 1     
                   
               
               
                 376703 
                 7.46 
                 93 
                   
                 1     
                   
                   
                 0     
                 0.164756 
                 0.835244 
                   
                 0.437086 
                 0.562914 
               
               
                 376709 
                  8.439 
                 92 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.43985  
                 0.56015  
               
               
                 376715 
                  2.451 
                 98 
                   
                 0.4    
                 0.6    
                   
                 0     
                 1     
                   
                   
                 0.285124 
                 0.714876 
               
               
                 376721 
                  5.544 
                 94.456 
                   
                 0.85   
                   
                   
                 0.15   
                 0.197595 
                 0.802405 
                   
                 0.327751 
                 0.672249 
               
               
                 376725 
                  7.766 
                 71.445 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.413333 
                 0.586667 
               
               
                 376729 
                  4.728 
                 95 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.414343 
                 0.585657 
               
               
                 376733 
                  9.277 
                 91 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.474359 
                 0.525641 
               
               
                 376739 
                 11.358 
                 70.083 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.719481 
                 0.280519 
               
               
                 376745 
                 18.482 
                 82 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.479675 
                 0.520325 
               
               
                 376750 
                 5.36 
                 94.64 
                   
                 1     
                   
                   
                 0     
                 0.153509 
                 0.846491 
                   
                 1     
                   
               
               
                 376755 
                  4.451 
                 96 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.509225 
                 0.490775 
               
               
                 376760 
                  6.609 
                 93.391 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376765 
                  4.751 
                 57.868 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.305136 
                 0.694864 
               
               
                 376770 
                  7.176 
                 50.33 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 376775 
                 36    
                 63.705 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.527027 
                 0.472973 
               
               
                 376779 
                 5.25 
                 94.75 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.416107 
                 0.583893 
               
               
                 376785 
                 19.124 
                 80.876 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.443333 
                 0.556667 
               
               
                 376789 
                  2.556 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.385294 
                 0.614706 
               
               
                 376793 
                  9.161 
                 79.135 
                   
                 1     
                   
                   
                 0     
                 0.120112 
                 0.879888 
                   
                 1     
                   
               
               
                 376797 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376801 
                 16.037 
                 83.963 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376806 
                  3.911 
                 96 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376811 
                 35.57  
                 41.522 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 376817 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376822 
                 3.67 
                 96 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.304688 
                 0.695313 
               
               
                 376826 
                  1.521 
                 98.479 
                   
                 1     
                   
                   
                 0     
                 0.097792 
                 0.902208 
                   
                 0.487738 
                 0.512262 
               
               
                 376830 
                  3.377 
                 14.015 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376834 
                  3.397 
                 96.613 
                   
                 1     
                   
                   
                 0     
                 0.495667 
                 0.504333 
                   
                 0.229858 
                 0.770142 
               
               
                 376838 
                  2.644 
                 97.356 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.25463  
                 0.74537  
               
               
                 376842 
                  2.839 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.149877 
                 0.850123 
               
               
                 376846 
                 2.28 
                 97.72 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376850 
                 2.28 
                 97.72 
                   
                 1     
                   
                   
                 0     
                 0.332192 
                 0.667808 
                   
                 0.735577 
                 0.264423 
               
               
                 376854 
                  0.963 
                 99.037 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.533951 
                 0.466049 
               
               
                 376858 
                  2.903 
                 97.097 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.513889 
                 0.486111 
               
               
                 376862 
                  2.567 
                 97.433 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.666667 
                 0.333333 
               
               
                 376867 
                  2.062 
                 97.938 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.495702 
                 0.504298 
               
               
                 376872 
                 50.192 
                 50 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.171254 
                 0.828746 
               
               
                 376879 
                 10.705 
                 65.812 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.303207 
                 0.696793 
               
               
                 376885 
                  1.202 
                 77.072 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.232628 
                 0.767372 
               
               
                 376889 
                  9.579 
                 83.982 
                   
                   
                   
                   
                   
                   
                   
                   
                 0.546   
                 0.454   
               
               
                 376897 
                  6.197 
                 93.803 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.540146 
                 0.459854 
               
               
                 376902 
                  7.841 
                 92.1259 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 376907 
                  2.508 
                 97.492 
                   
                 0.81   
                   
                   
                 0.19   
                 0.213946 
                 0.786054 
                   
                 0.176152 
                 0.823848 
               
               
                 376913 
                 1.59 
                 98.41 
                   
                 0.63763  
                   
                   
                 0.3621  
                 0.589091 
                 0.410909 
                   
                 0.142539 
                 0.857461 
               
               
                 376918 
                  2.689 
                 97 
                   
                 0.607   
                   
                   
                 0.393   
                 1     
                   
                   
                 0.310078 
                 0.689922 
               
               
                 376925 
                  2.701 
                 97.479 
                   
                   
                   
                   
                 1     
                 1     
                   
                   
                 0.245283 
                 0.754717 
               
               
                 376931 
                  3.825 
                 17 
                 23.971 
                 0.845   
                   
                   
                 0.15   
                 1     
                   
                   
                 1     
                   
               
               
                 376937 
                  6.425 
                 73 
                  6.777 
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 376941 
                 6.09 
                 93.91 
                   
                 0.995   
                   
                   
                 0     
                 0.104452 
                 0.895548 
                   
                 0.456731 
                 0.543269 
               
               
                 376949 
                  4.915 
                 95.085 
                   
                 0.136   
                 0.805   
                   
                 0.059   
                 1     
                   
                   
                 0.432161 
                 0.567839 
               
               
                 376955 
                  3.589 
                 96.411 
                   
                 0.197   
                 0.287   
                 0.51 
                 0     
                 1     
                   
                   
                 0.222841 
                 0.777159 
               
               
                 376962 
                  5.594 
                 86.48 
                  7.926 
                 0.636074 
                 0.636074 
                   
                 0.363926 
                 1     
                   
                   
                 0.321678 
                 0.678322 
               
               
                 376967 
                  6.547 
                 93.453 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.486869 
                 0.513131 
               
               
                 376972 
                  3.031 
                 96.969 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.42549  
                 0.57451  
               
               
                 376976 
                  3.387 
                 96.613 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.487965 
                 0.512035 
               
               
                 376983 
                 34.082 
                 66 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 376991 
                  9.467 
                 90.533 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.501219 
                 0.498781 
               
               
                 376995 
                 0   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377001 
                  2.545 
                 97.455 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.519288 
                 0.480712 
               
               
                 377006 
                 0   
                 66.136 
                 33.864 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377011 
                  2.172 
                 79 
                   
                   
                   
                   
                   
                 0.544944 
                 0.455056 
                   
                 0.247312 
                 0.752688 
               
               
                 377015 
                  9.233 
                 57 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                 0     
               
               
                 377020 
                  7.127 
                 85 
                 51.551 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377025 
                  8.132 
                 85 
                 15.948 
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377031 
                  3.924 
                 91 
                   
                   
                   
                   
                   
                 0.473149 
                 0.526851 
                   
                 0.577419 
                 0.422581 
               
               
                 377036 
                  3.788 
                 96 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377041 
                 16.2361 
                 83.982 
                   
                 1     
                   
                   
                 0     
                 0.315871 
                 0.684129 
                   
                 1     
                   
               
               
                 377045 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377050 
                 11.164 
                 77 
                 10.496 
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 377056 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377060 
                  7.075 
                 77 
                 33.064 
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377064 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377068 
                  3.292 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.299145 
                 0.700855 
               
               
                 377073 
                  1.927 
                 98.074 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.376947 
                 0.623053 
               
               
                 377077 
                  2.583 
                 97 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.110749 
                 0.889251 
               
               
                 377082 
                  1.135 
                 99 
                   
                 1     
                   
                   
                 0     
                 0.225086 
                 0.774914 
                   
                 0.071429 
                 0.928571 
               
               
                 377087 
                  2.006 
                 97.929 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.274933 
                 0.725067 
               
               
                 377092 
                  1.994 
                 98.006 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.094395 
                 0.905605 
               
               
                 377097 
                  1.261 
                 98.739 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 377102 
                 2.11 
                 97.767 
                   
                 1     
                   
                   
                 0     
                 0.164251 
                 0.835749 
                   
                 0.501319 
                 0.498681 
               
               
                 377107 
                  1.501 
                 98 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.485893 
                 0.514107 
               
               
                 377112 
                  3.322 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.468657 
                 0.531343 
               
               
                 377116 
                 2.65 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.557003 
                 0.442997 
               
               
                 377121 
                  3.284 
                 97 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.459701 
                 0.540299 
               
               
                 377126 
                 12.404 
                 66 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377133 
                  6.553 
                 57 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.436508 
                 0.563492 
               
               
                 377138 
                  3.988 
                 73 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.400932 
                 0.599068 
               
               
                 377142 
                  2.077 
                 98 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.52861  
                 0.47139  
               
               
                 377148 
                  3.772 
                 96 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.529745 
                 0.470255 
               
               
                 377152 
                  1.436 
                 99 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.50289  
                 0.49711  
               
               
                 377156 
                  2.614 
                 77 
                   
                 1     
                   
                   
                 0     
                 0.118734 
                 0.886544 
                   
                 1     
                   
               
               
                 377162 
                  1.394 
                 98.536 
                   
                 0.64   
                   
                   
                 0.33   
                 0.065466 
                 0.145663 
                 0.485   
                 0.172237 
                 0.827763 
               
               
                 377166 
                  5.575 
                 94 
                  2.313 
                 0.295   
                 0.45   
                   
                 0.254   
                 1     
                   
                   
                 0.359116 
                 0.640884 
               
               
                 377172 
                  3.898 
                 96 
                   
                 0.261   
                 0.325   
                   
                 0.414   
                 1     
                   
                   
                 0.349333 
                 0.650667 
               
               
                 377176 
                 19.399 
                 77 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 377182 
                 32.588 
                 50 
                   
                   
                   
                   
                 1     
                 1     
                   
                   
                 1     
                   
               
               
                 377186 
                  6.813 
                 93 
                   
                 1     
                   
                   
                 0     
                 0.171975 
                 0.828025 
                   
                 0.454054 
                 0.545946 
               
               
                 377193 
                  1.394 
                 98.605 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.446108 
                 0.553892 
               
               
                 377200 
                  5.103 
                 95 
                   
                 0.15   
                 0.165   
                 0.688 
                 0     
                 1     
                   
                   
                 0.334448 
                 0.665552 
               
               
                 377208 
                  3.092 
                 97 
                   
                 0.84   
                   
                   
                 0.16   
                 0.229787 
                 0.770213 
                   
                 0.338983 
                 0.661017 
               
               
                 377212 
                 20.005 
                 80 
                   
                 0.867   
                   
                   
                 0.133   
                 1     
                   
                   
                 0.412888 
                 0.587112 
               
               
                 377217 
                  5.089 
                 95 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.44709  
                 0.55291  
               
               
                 377223 
                 7.45 
                 93 
                   
                 1     
                   
                   
                 0     
                 0.065574 
                 0.934426 
                   
                 0.497423 
                 0.502577 
               
               
                 377229 
                 20.138 
                 62 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377236 
                 18.44  
                 82 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.488439 
                 0.511561 
               
               
                 377241 
                 10.022 
                 90 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377246 
                 7   
                 92.694 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.519878 
                 0.480122 
               
               
                 377252 
                 16    
                 83.804 
                  6.111 
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377256 
                 0   
                 100 
                   
                   
                   
                   
                   
                   
                   
                   
                 1     
                   
               
               
                 377261 
                 0   
                 100 
                   
                 1     
                   
                   
                 0     
                   
                   
                   
                 1     
                   
               
               
                 377266 
                 0   
                 100 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 1     
                   
               
               
                 377272 
                  8.675 
                 84 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.387841 
                 0.612159 
               
               
                 377276 
                 11.006 
                 88.626 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.486486 
                 0.513514 
               
               
                 377281 
                  4.722 
                 95.279 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 0.485488 
                 0.514512 
               
               
                 377286 
                  5.497 
                 64 
                   
                 1     
                   
                   
                 0     
                 0.134328 
                 0.865672 
                   
                 1     
                   
               
               
                 377290 
                 0   
                 100 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 377295 
                 13.807 
                 10.899 
                   
                 1     
                   
                   
                 0     
                 1     
                   
                   
                 1     
                   
               
               
                 377300 
                  6.932 
                 87 
                   
                   
                   
                   
                   
                 1     
                   
                   
                 0.566553 
                 0.433447 
               
               
                 377305 
                 11.808 
                 83 
                   
                 1     
                   
                   
                 0     
                 0.331418 
                 0.668582 
                   
                 0.8125  
                 0.1875  
               
               
                 377310 
                 0   
                 100 
               
               
                   
               
            
           
         
       
     
     All publications, patents, and patent applications discussed and cited herein are hereby incorporated by reference in their entireties. It is understood that the disclosed invention is not limited to the particular methodology, protocols and materials described as these can vary. It is also understood that the terminology used herein is for the purposes of describing particular embodiments only and is not intended to limit the scope of the appended claims. 
     Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the claims that follow. 
     ABBREVIATIONS 
     Abbreviated terms used throughout this specification are defined as follows.
     aa, AA amino acid   AEI allelic expression imbalance   ANOVA analysis of variance   BSA bovine serum albumin   CDR complementarity determining regions   CHO Chinese hamster ovary cells   CPM charge pair mutation   DMEM Dulbecco&#39;s Modified Eagle Medium   DMSO dimethyl sulfoxide   ELISA enzyme-linked immunosorbent assay   eQTL expression quantitative trait loci   ESI-TOF electrospray ionization time of flight   ESN exhausted supernatant   FACS fluorescence-activated cell sorting   FBS fetal bovine serum   FPLC fast protein liquid chromatography   FVB a strain of mice inbred for the Friend leukemia virus 1b (Fv1b) allele   H&amp;E Hematoxylin and eosin   HA hypoxanthine   HIC hydrophobic interaction chromatography   HPLC high performance liquid chromatography   HRP horse radish peroxidase   HUVEC human umbilical vein epithelial cell   IBD inflammatory bowel disease   IDMEM DMEM without glutamine   IFN interferon   IL interleukin   MCP monocyte chemotactic protein   MSD macromolecular structure database   NA nucleic acid   PBMC peripheral blood mononuclear cell   PBS phosphate-buffered saline   PCR polymerase chain reaction   PEG polyethylene glycol   PEI polyethylenimine   QTL quantitative trait loci   RPMI media developed at Roswell Park Memorial Institute   RT-PCR polymerase chain reaction at room temperature   SNP single nucleotide polymorphism   TFA trifluoroacetic acid   TL1A TNF-like ligand 1A (TNFSF15)   TMB tetramethylbenzene   TNF tumor necrosis factor-α