Abstract:
This invention relates to specific antibodies against ganglioside GD3 called MB3.6 and against protein prostate specific membrane antigen (PSMA) called 3D8, 4D4 and 3E11 when prepared as chimeric molecules with signaling molecules of T cells and other effector cells, and the use thereof in the treatment of cancers expressing these antigens.

Description:
STATEMENT ON FEDERALLY-SPONSORED R&amp;D  
         [0001]    No federal funds were used in the creation of this invention.  
         BACKGROUND OF THE INVENTION  
         [0002]    More than 500,000 Americans die each year from cancers that have proven refractory to traditional methods of treatment, for which new strategies are urgently required. Tumor-associated antigens are selected for therapeutic targets based on their high expression on tumor tissue and a lower expression in normal tissues that will plausibly allow selective targeting of tumorous expression of the antigen. Ganglioside GD3 is expressed at high levels on melanoma, small cell lung cancer and other neuroendocrine tumors. Prostate-specific membrane antigen (PSMA) is selectively expressed at high levels in prostate cancer and other tumors. T cells can penetrate virtually every biologic space and have the power to dispose of normal or malignant cells as seen in viral and autoimmune diseases and in the rare spontaneous remissions of cancer. However, T cells are readily tolerant to self or tumor antigens, and “immune surveillance” has manifestly failed in every cancer that is clinically apparent. There is a strong need and value for means to direct T cells against GD3-expressing and PSMA-expressing cancers. This patent describes specific molecules and means to achieve this goal.  
         BRIEF SUMMARY OF THE INVENTION  
         [0003]    An antibody against GD3 has been prepared called MB3.6. GD3 is expressed at high levels on melanoma and other neuroendocrine tumors, and low levels on normal tissues. Antibodies against PSMA have been prepared called 3D8, 4D4, 3E11. PSMA is expressed principally in prostate tissue, a non-essential organ, and in prostate cancers. It is the goal of this patent to supply the specificities and affinities to patient T cells without regard for their “endogenous” T cell receptor repertoire, directed by antibody-defined recognition to kill malignant cells based on their expression of antigen. This is achieved by preparing chimeric molecules of these specific antibodies with molecules derived from T cells or related effector cell molecules, which will redirect T cells or other effector cells against the tumor cells in a focused anti-tumor immune response by “re-educating” the patient&#39;s immune system. 
       
    
    
     BRIEF DESCRIPTION OF DRAWINGS  
       [0004]    [0004]FIG. 1 shows a chimeric antibody-T cell receptor that employs the zeta chain of the TCR. In this example, a single chain Fv (sFv) version of hMN14 is linked by a CD8α hinge to the TCR zeta chain. The CD8α hinge has been further modified to remove the cysteines involved in CD8 dimerization to improve surface expression.  
         [0005]    [0005]FIG. 2A shows the near absence of heterodimer molecules when the native CD8α hinge is employed, although it would be predicted to be the dominant species, with a lower net expression of chimeric molecule relative to endogenous zeta chain. FIG. 2B shows the effect of removing the cysteines, which now allows much increased net expression of chimeric molecule when heterodimer can be expressed.  
         [0006]    [0006]FIG. 3 shows diagram and DNA sequence of a chimeric sFv IgTCR, including the CD8α hinge modified-to-remove cysteines, within a retroviral vector. This example IgTCR molecule (using hMN14 antibody specific to CEA antigen, not part of this application) occupies nucleotides 2426 to 3766. (The vector sequences are incidental.) Equivalent versions using the antibodies MB3.6, 3D8, 4D4, 3E11 are prepared in analogous manner to create IgTCR, or other Ig-chimeric molecules.  
         [0007]    [0007]FIG. 4 shows the DNA sequence of:  
         [0008]    A., B. leader plus VH and leader plus VL that specifies MB3.6.  
         [0009]    C. As example, the VL and leader are joined with linker to VH to create MB3.6 sFv as shown, that is subsequently used in creating chimeric molecules. Other means of generating sFv are possible and included under this claim, as well as other means of creating antibody chimeric molecules under the intent of this invention.  
         [0010]    D., E. leader plus VH and leader plus VL that specifies 3D8 (includes C domain sequences).  
         [0011]    F., G. leader plus VH and leader plus VL that specifies 4D4 (includes C domain sequences).  
         [0012]    H., I. leader plus VH and leader plus VL that specifies 3E11 (includes C domain sequences).  
         [0013]    These sequences are modified to prepare the sFv used in FIG. 1 and FIG. 3, and similarly for other constructs.  
         [0014]    [0014]FIG. 5 shows example of the effect of MB3.6 IgTCR-modified T cells in killing GD3-positive tumor cells, but sparing GD3-negative cells. Other examples with 3D8, 4D4, 3E11 would show specific killing against PSMA-positive cells but not against PSMA-negative cells.  
         [0015]    [0015]FIG. 6 shows example of the effect of IgTCR (signal 1) using hMN14 antibody chimerics against CEA (not part of this application) on causing sustained CEA+ tumor cell killing when stimulated in conjunction with CD28 (signal 2) stimulation of the gene-modified T cells via B7 antigen expressed in the tumor cells. (A) Signal 1 alone from tumor cells leads to AICD with declining effector cell numbers, that is reversed with signal 1+2. (B) Signal 1 leads to limited duration of tumor killing because of declining T cell numbers. (C) Signal 1+2 leads to sustained tumor killing because of the sustained and expanding T cell numbers. This example justifies design of IgCD28 molecules to modify patient T cells to supply the second signal on contact with antigen that is necessary to suppress effector cell death and achieve sustained killing activity. The intent of this example is the expectation of utility with analogous constructs using the Ig sequences of this application.  
         [0016]    [0016]FIG. 7 shows an example of a design for an IgCD28 using MB3.6, 3D8, 4D4, 3E11. This also uses a modified CD8α hinge. Similar designs for other chimeric molecules with these antibodies are envisioned, with or without hinge that is the same or different. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0017]    This patent is intended to cover all chimeric molecules created with the specified antibodies (Ig) (MB3.6, 3D8, 4D4, 3E11) (defined by the variable region sequences of FIG. 4) or their derivatives with cell surface molecules which could be used in redirecting and/or activating T cells or other effector cells in the recognition and attack against tumors expressing the antigens recognized by these antibodies. Other specific antibodies which the inventor or his agents obtain with rights will be similarly appended as claims at such future appropriate time. The chimeric molecules of this claim include, but are not limited to, the following molecules: IgTCR (FIGS. 1&amp;3), which has an antibody binding domain from these antibodies fused to one or more chains of the T cell receptor complex; IgCD28 (FIG. 7), which has an antibody binding domain from these antibodies fused to the CD28 T cell co-receptor molecule; IgLFA-1, which has an antibody binding domain from these antibodies fused to the LFA-1 T cell co-receptor/adhesion molecule; IgCD2, which has an antibody binding domain from these specific antibodies fused to the CD2 T cell co-receptor/adhesion molecule; and by analogy, any other T cell or effector cell molecules which are usefully employed in chimeric structures with these antibody binding domains. The chimeric molecules may themselves incorporate cytoplasmic signaling domains, as in the previous examples. Or the chimeric molecules may instead be non-signaling, such as examples of Ig linked to TCR α or β chains, or Ig linked to Fc receptor (FcR) non-signaling chains, that in turn associate with signaling chains to activate cellular functions. These chimeric molecules may additionally incorporate spacer domains or epitope tags. Single-chain Fv (sFv) versions of these antibodies have been favored for use in these constructs, but Fab or other IgG chimeric molecules would be equally included under this invention. The initial description of some of these preparations is contained in Yun et al, 2000. This demonstrates reduction to practice of the concepts contained herein for the MB3.6 antibody, with expectation of similar results for the other antibodies specified in this invention.  
         [0018]    The invention additionally allows for the presence of a (GSGGS)3 linker in the sFv of the Ig portion of the chimeric molecules (e.g., FIG. 4C). Whereas the sFv antibodies may frequently not fold properly to maintain stability, I included the extra serine to improve hydration and sFv folding versus the typical (GGGGS)3 linker that has been associated in some cases with abolished or diminished sFv affinity (e.g., Brinkmann et al, 1993). This strategy with an antibody not covered under this patent (hMN14) led to an sFv virtually indistinguishable from the monovalent binding affinity of the parental antibody (Nolan et al, 1999). Such tests have not been performed with the current antibodies, but all have maintained antigen recognition after sFv modification with this linker.  
         [0019]    The invention additionally allows for the modification-to-remove cysteines in the CD8α hinge domain to improve the surface expression of the chimeric molecules (Nolan et al, 1999). Free cysteines of the hinge of the heterodimer of zeta:sFv-hinge-zeta target this molecular complex for destruction, reducing the net amount of chimeric molecule expression on the cell surface. (The homodimer (sFv-hinge-zeta) 2  has safe pairing of cysteines to spare this specific configuration from destruction. More heterodimer is expected because of binomial considerations where the endogenous zeta exceeds the transduced zeta chimera as is typical.) This principle is demonstrated by the poor expression of heterodimers of such molecules where the cysteine residues are retained (Moritz et al, 1995) and their excellent expression when I modified-to-remove these cysteines (Nolan et al, 1999) (FIG. 2). The efficacy of T cell functions through surface receptors are generally higher with higher surface expression, which the rescue (i.e., non-destruction) of heterodimers would allow. These chimeric molecules are introduced into patient T cells by gene therapy techniques, such as by retroviral vector transduction or other methods. This method of improving cell surface expression is cross-referenced (Junghans Provisional Patent No. 60/250,087).  
         [0020]    In one example, IgTCR (FIG. 1) provides signal 1, which directs T cell killing (e.g., FIG. 5); IgCD28 (FIG. 7) provides signal 2, which suppresses activation induced cell death of T cells and allows sustained proliferation and survival (FIG. 6); and IgLFA1, which provides signal 3 and supports secretion of interleukin 2, an essential T cell growth factor. Combinations of signals can yield improved T cell survival and tumor cell killing (FIG. 6). The invention allows for use of these and/or analogous chimeric molecules of hMN14 alone or in any combination.  
         [0021]    The combination use of such chimeric molecules in treatment of cancers is a further part of the claim. This applies an understanding that more than one signal is required for sustained antitumor efficacy. This application specifically envisions that the same antibody binding domain is applied in the additional chimeric receptor molecules such that encounter with the same tumor antigen successfully triggers more than one signal in the effector cell. Alternatively, additional signaling chimeric molecules may have engineered Ig specificities which direct them to different surface molecules on the tumor cell, rather than to the same one, to avoid binding site competition or to regulate the amount of receptor stimulation where this regulation enhances the desired outcome of antitumor efficacy in therapy.  
         [0022]    The purpose of this invention is to educate immune effector cells to attack GD3-expressing or PSMA-expressing tumor cells. Advantages are that the sequences used to recognize GD3 or PSMA in their conjugation with T cell molecules leads to direct recognition of GD3+ or PSMA+ tumors by human T cells, and hinge and sFv linker modifications make the surface expression more efficient with advantages in anti-tumor activity. Presently, treatments for these cancers are chemotherapy, immunotherapy, surgery and radiation, which are rarely or never curative for metastatic disease. A critical component of this patent for therapy is the specific antibodies that recognize these antigens. No other IgTCR or Ig-T cell molecules has the amino acid sequence of the GD3 or PSMA antibody recognition domains specified herein, and which I have proven to be effective (e.g., FIG. 5). There is no patent of these sequences in chimeric state with T cell or other effector cell molecules, or with the use of a modified hinge structure. This purpose is expanded to other tumor-associated antigens as appropriate to other antibodies as obtained with rights by the inventor or his agents, which will be appended as supplemental claims at such appropriate time.  
         [0023]    An invention exists as to the general chimeric Ig molecules with cell receptor proteins (Capon et al, 1995). This invention is distinguished by the uniqueness of the antibody sequences employed, by the new concept in modification of hinge domains that improves the expression in cells, and by the combination of such chimeric receptor molecules expressed in effector cells which are stimulated in concert specifically by the same tumor antigen or by a different tumor antigen or antigens. The sequences of these antibodies were not previously patented. The claims of the present patent are restricted to the use of these antibodies and sequences in the preparation of these chimeric molecules for the purposes herein described. These claims as pertaining to these sequences do not extend to other potential uses of these antibodies and their derivatives, which are reserved for potential future applications.  
     
       
       
         1 
         
           
             19  
           
           
             1  
             7654  
             DNA  
             Homo sapiens and Mus sp.  
             
               CDS  
               (2428)..(3759)  
               Chimeric IgTCR sequence contained in retroviral 
      vector.  Retroviral vector sequence (non-coding regions) are 
      incidental to the invention.  The translated (coding region) is 
      relevant to the invention. (pertinent to Figure 3.)  
             
           
            1 

aagcttgcat gcctgcaggt cgactctagg cacataaaga aaaacataac taaccaagct     60 

gcagccgaga cagtgaaaag aaccgttaaa acggtttgtt ttaaataaac tgaattattt    120 

agagtcattt ctttggtagg aaagtacatt ggcacgtaaa ggagcccaaa gcaatctgtg    180 

gaaagcccag gctgggagcc cagcagtttg catcccctcc tggcgtgtac ctaagggttt    240 

cttaattgtg tggtttctaa atcttccaga gggtttgtct cattcacttc cacttcggtg    300 

cacaatactt ggacgcggat ttactgtctt agcatctatc ggtggccctt cgattgaggc    360 

tgaacctgag gcccacttct tcagcttgtt aaggagagca caagcaccag aagaggctga    420 

cccggcagac ctgtgggcat ttttaacaag ggcctcctgg gtctgtggga ggcaggctta    480 

cataaggtgc aaattagaaa tataaataat aagcccatat caatttgtca tcttttttta    540 

agctcaagtt ttgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt    600 

ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca    660 

gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccgctc    720 

agggccaaga acagttggaa caggagaata tgggccaaac aggatatctg tggtaagcag    780 

ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca    840 

gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc    900 

cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg    960 

agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt   1020 

cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc   1080 

gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt   1140 

tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga   1200 

ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt   1260 

tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg   1320 

aactgacgag ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg acttcggggg   1380 

ccgtttttgt ggcccgacct gagtcctaaa atcccgatcg tttaggactc tttggtgcac   1440 

cccccttaga ggagggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc   1500 

ctccgtctga atttttgctt tcggtttggg accgaagccg cgccgcgcgt cttgtctgct   1560 

gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaatatgg   1620 

gcccgggcta gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg   1680 

agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct   1740 

ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc   1800 

tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg   1860 

tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct   1920 

ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac   1980 

ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg   2040 

cccccatatg gccatatgag atcttatatg gggcaccccc gccccttgta aacttccctg   2100 

accctgacat gacaagagtt actaacagcc cctctctcca agctcactta caggcttcta   2160 

cttagtccag cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg   2220 

accggtggta cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac   2280 

taagaaccta gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc   2340 

cctcaaagta gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc   2400 

cgggggtgga ccatcctcta gactgcc atg gga tgg agc tgt atc atc ctc ttc   2454 
                              Met Gly Trp Ser Cys Ile Ile Leu Phe 
                              1               5 

ttg gta gca aca gct aca ggt gtc cac tcc gac atc cag ctg acc cag     2502 
Leu Val Ala Thr Ala Thr Gly Val His Ser Asp Ile Gln Leu Thr Gln 
10                  15                  20                  25 

agc cca agc agc ctg agc gcc agc gtg ggt gac aga gtg acc atc acc     2550 
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 
                30                  35                  40 

tgt aag gcc agt cag gat gtg ggt act tct gta gct tgg tac cag cag     2598 
Cys Lys Ala Ser Gln Asp Val Gly Thr Ser Val Ala Trp Tyr Gln Gln 
            45                  50                  55 

aag cca ggt aag gct cca aag ctg ctg atc tac tgg aca tcc acc cgg     2646 
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Trp Thr Ser Thr Arg 
        60                  65                  70 

cac act ggt gtg cca agc aga ttc agc ggt agc ggt agc ggt acc gac     2694 
His Thr Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 
    75                  80                  85 

ttc acc ttc acc atc agc agc ctc cag cca gag gac atc gcc acc tac     2742 
Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr 
90                  95                  100                 105 

tac tgc cag caa tat agc ctc tat cgg tcg ttc ggc caa ggg acc aag     2790 
Tyr Cys Gln Gln Tyr Ser Leu Tyr Arg Ser Phe Gly Gln Gly Thr Lys 
                110                 115                 120 

gtg gaa atc aaa cga ggt ggc tca gga tcg ggt gga tcc ggc tct ggt     2838 
Val Glu Ile Lys Arg Gly Gly Ser Gly Ser Gly Gly Ser Gly Ser Gly 
            125                 130                 135 

ggc tca gga tcg gag gtc caa ctg gtg gag agc ggt gga ggt gtt gtg     2886 
Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val 
        140                 145                 150 

caa cct ggc cgg tcc ctg cgc ctg tcc tgc tcc gca tct ggc ttc gat     2934 
Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Asp 
    155                 160                 165 

ttc acc aca tat tgg atg agt tgg gtg aga cag gca cct gga aaa ggt     2982 
Phe Thr Thr Tyr Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 
170                 175                 180                 185 

ctt gag tgg att gga gaa att cat cca gat agc agt acg att aac tat     3030 
Leu Glu Trp Ile Gly Glu Ile His Pro Asp Ser Ser Thr Ile Asn Tyr 
                190                 195                 200 

gcg ccg tct cta aag gat aga ttt aca ata tcg cga gac aac gcc aag     3078 
Ala Pro Ser Leu Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 
            205                 210                 215 

aac aca ttg ttc ctg caa atg gac agc ctg aga ccc gaa gac acc ggg     3126 
Asn Thr Leu Phe Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Gly 
        220                 225                 230 

gtc tat ttt tgt gca agc ctt tac ttc ggc ttc ccc tgg ttt gct tat     3174 
Val Tyr Phe Cys Ala Ser Leu Tyr Phe Gly Phe Pro Trp Phe Ala Tyr 
    235                 240                 245 

tgg ggc caa ggg acc ccg gtc acc gtc tcc agt gct aag ccc acc acg     3222 
Trp Gly Gln Gly Thr Pro Val Thr Val Ser Ser Ala Lys Pro Thr Thr 
250                 255                 260                 265 

acg cca gcg ccg cga cca cca aca ccg gcg ccc acc atc gcg tcg cag     3270 
Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln 
                270                 275                 280 

ccc ctg tcc ctg cgc cca gag gcg gct cgg cca gcg gcg ggg ggc gca     3318 
Pro Leu Ser Leu Arg Pro Glu Ala Ala Arg Pro Ala Ala Gly Gly Ala 
            285                 290                 295 

gtg cac acg agg ggg ctg gac ttc gcc ctg gat ccc aaa ctc tgc tac     3366 
Val His Thr Arg Gly Leu Asp Phe Ala Leu Asp Pro Lys Leu Cys Tyr 
        300                 305                 310 

ctg ctg gat gga atc ctc ttc atc tat ggt gtc att ctc act gcc ttg     3414 
Leu Leu Asp Gly Ile Leu Phe Ile Tyr Gly Val Ile Leu Thr Ala Leu 
    315                 320                 325 

ttc ctg aga gtg aag ttc agc agg agc gca gag ccc ccc gcg tac cag     3462 
Phe Leu Arg Val Lys Phe Ser Arg Ser Ala Glu Pro Pro Ala Tyr Gln 
330                 335                 340                 345 

cag ggc cag aac cag ctc tat aac gag ctc aat cta gga cga aga gag     3510 
Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 
                350                 355                 360 

gag tac gat gtt ttg gac aag aga cgt ggc cgg gac cct gag atg ggg     3558 
Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 
            365                 370                 375 

gga aag ccg aga agg aag aac cct cag gaa ggc ctg tac aat gaa ctg     3606 
Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu 
        380                 385                 390 

cag aaa gat aag atg gcg gag gcc tac agt gag att ggg atg aaa ggc     3654 
Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly 
    395                 400                 405 

gag cgc cgg agg ggc aag ggg cac gat ggc ctt tac cag ggt ctc agt     3702 
Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser 
410                 415                 420                 425 

aca gcc acc aag gac acc tac gac gcc ctt cac atg cag gcc ctg ccc     3750 
Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro 
                430                 435                 440 

cct cgc taa ctcgacgcgg ccgcggatcc ggattagtcc aatttgttaa             3799 
Pro Arg 

agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc   3859 

tatagagtac gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg   3919 

aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc   3979 

atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag   4039 

cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccgct cagggccaag   4099 

aacagttgga acaggagaat atgggccaaa caggatatct gtggtaagca gttcctgccc   4159 

cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga   4219 

gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga   4279 

actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata   4339 

aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt   4399 

acccgtgttc tcaataaacc ctcttgcagt tgcatccgac tcgtggtctc gctgttcctt   4459 

gggagggtct ctctgagtga ttgactaccc gtcagcgggg tctttcagtt tctcccacct   4519 

acacaggtct cactaacatt cctgatgtgc cgcagggact ccgtcagccc ggtttttgtt   4579 

tataataaaa tgcaagaaca gtgttccctt caagccagac tacatcctga ctctcggctt   4639 

tataaaagaa tgttgaaggg ctctgtggac tatctgccac acgacttttt aagattttta   4699 

tgcctcctgg atgagggatt tagtcaatct atcctcgtct attttgctgg cttctccgta   4759 

ttttaaattt ctagtttgca ctcccttcct gagagcacgg cgattgcaga gtagttaata   4819 

ctctgagggc aggcttctgt gaaaaggttg cctgggctca gtgtgagatt ttgccataaa   4879 

aaggggtcct gcccctgtgt acagacagat cggaatctag agtgcatact cagagtcccc   4939 

gcggttccgg ggctctgatc tcagggcatc tttgcctaga gatcctctac gccggacgca   4999 

tcgtggccgg gtaccgagct cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa   5059 

ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg   5119 

gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca   5179 

gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg   5239 

tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg   5299 

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg   5359 

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa   5419 

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg   5479 

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc   5539 

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc   5599 

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc   5659 

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg   5719 

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc   5779 

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga   5839 

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc   5899 

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac   5959 

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg   6019 

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc   6079 

acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa   6139 

ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta   6199 

ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt   6259 

tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag   6319 

tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca   6379 

gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc   6439 

tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt   6499 

tgttgccatt gctacaggct cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc   6559 

tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt   6619 

agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg   6679 

gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg   6739 

actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct   6799 

tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc   6859 

attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt   6919 

tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt   6979 

tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg   7039 

aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat   7099 

tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg   7159 

cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta   7219 

acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt   7279 

gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc   7339 

gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt   7399 

aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg   7459 

cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac   7519 

tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga   7579 

tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa   7639 

acgacggcca gtgcc                                                    7654 

 
           
             2  
             443  
             PRT  
             Homo sapiens and Mus sp.  
           
            2 

Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 
1               5                   10                  15 

Val His Ser Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala 
            20                  25                  30 

Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val 
        35                  40                  45 

Gly Thr Ser Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys 
    50                  55                  60 

Leu Leu Ile Tyr Trp Thr Ser Thr Arg His Thr Gly Val Pro Ser Arg 
65                  70                  75                  80 

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser 
                85                  90                  95 

Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Leu 
            100                 105                 110 

Tyr Arg Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Gly Gly 
        115                 120                 125 

Ser Gly Ser Gly Gly Ser Gly Ser Gly Gly Ser Gly Ser Glu Val Gln 
    130                 135                 140 

Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Arg 
145                 150                 155                 160 

Leu Ser Cys Ser Ala Ser Gly Phe Asp Phe Thr Thr Tyr Trp Met Ser 
                165                 170                 175 

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile 
            180                 185                 190 

His Pro Asp Ser Ser Thr Ile Asn Tyr Ala Pro Ser Leu Lys Asp Arg 
        195                 200                 205 

Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met 
    210                 215                 220 

Asp Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Phe Cys Ala Ser Leu 
225                 230                 235                 240 

Tyr Phe Gly Phe Pro Trp Phe Ala Tyr Trp Gly Gln Gly Thr Pro Val 
                245                 250                 255 

Thr Val Ser Ser Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro 
            260                 265                 270 

Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 
        275                 280                 285 

Ala Ala Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 
    290                 295                 300 

Phe Ala Leu Asp Pro Lys Leu Cys Tyr Leu Leu Asp Gly Ile Leu Phe 
305                 310                 315                 320 

Ile Tyr Gly Val Ile Leu Thr Ala Leu Phe Leu Arg Val Lys Phe Ser 
                325                 330                 335 

Arg Ser Ala Glu Pro Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr 
            340                 345                 350 

Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys 
        355                 360                 365 

Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn 
    370                 375                 380 

Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu 
385                 390                 395                 400 

Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly 
                405                 410                 415 

His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr 
            420                 425                 430 

Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
        435                 440 

 
           
             3  
             504  
             DNA  
             Mus sp.  
             
               CDS  
               (6)..(425)  
               MB3.6 Heavy chain V region, plus leader  
             
           
            3 

tcacc atg aac ttc ggg ttc agc ttg att ttc ctt gtc ctt gtt tta aaa     50 
      Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys 
      1               5                   10                  15 

ggt gtc cag tgt gaa gtg gtg gtg gtg gag tct ggg gga ggc ttc gtg       98 
Gly Val Gln Cys Glu Val Val Val Val Glu Ser Gly Gly Gly Phe Val 
                20                  25                  30 

aag cct gga ggg tcc ctg aaa ctc tcc tgt gca gcc gct gga ttc act      146 
Lys Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ala Gly Phe Thr 
            35                  40                  45 

ttc agt aga tat gcc atg tct tgg gtt cgc cag act ccg gag aag agg      194 
Phe Ser Arg Tyr Ala Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg 
        50                  55                  60 

ctg gag tgg gtc gca acc ata agt agt ggt ggt agt cac acc tac tat      242 
Leu Glu Trp Val Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr 
    65                  70                  75 

cca gac agt gtg aag ggg cga ttc acc atc tcc aga gac aat gcc aag      290 
Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 
80                  85                  90                  95 

aac acc ctg tac ctg caa atg agc agt ctg agg tct gag gac acg gcc      338 
Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala 
                100                 105                 110 

ata tat tac tgt gca aga ccg ggt tac gac agg ggg gcc tgg ttt ttc      386 
Ile Tyr Tyr Cys Ala Arg Pro Gly Tyr Asp Arg Gly Ala Trp Phe Phe 
            115                 120                 125 

gat gtc tgg ggc gca ggg acc acg gtc acc gtc tcc tca ggtaagtgtg       435 
Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 
        130                 135                 140 

tcagggtttc acaagaggga ctaaagacat gtcagctaat gtgtgactaa tggtaatgtc    495 

actaagctt                                                            504 

 
           
             4  
             140  
             PRT  
             Mus sp.  
           
            4 

Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly 
1               5                   10                  15 

Val Gln Cys Glu Val Val Val Val Glu Ser Gly Gly Gly Phe Val Lys 
            20                  25                  30 

Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ala Gly Phe Thr Phe 
        35                  40                  45 

Ser Arg Tyr Ala Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu 
    50                  55                  60 

Glu Trp Val Ala Thr Ile Ser Ser Gly Gly Ser His Thr Tyr Tyr Pro 
65                  70                  75                  80 

Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 
                85                  90                  95 

Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Ile 
            100                 105                 110 

Tyr Tyr Cys Ala Arg Pro Gly Tyr Asp Arg Gly Ala Trp Phe Phe Asp 
        115                 120                 125 

Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 
    130                 135                 140 

 
           
             5  
             483  
             DNA  
             Mus sp.  
             
               CDS  
               (16)..(399)  
               MB3.6 Light chain V region, plus leader  
             
           
            5 

agggaaagct cgaag atg gtt ttc aca cct cag ata ctt gga ctt atg ctt      51 
                 Met Val Phe Thr Pro Gln Ile Leu Gly Leu Met Leu 
                 1               5                   10 

ttt tgg att tca gcc tcc aga ggt gat att gtg cta act cag tct cca       99 
Phe Trp Ile Ser Ala Ser Arg Gly Asp Ile Val Leu Thr Gln Ser Pro 
        15                  20                  25 

gcc acc ctg tct gtg act cca gga gat agc gtc agt ctt tcc tgc agg      147 
Ala Thr Leu Ser Val Thr Pro Gly Asp Ser Val Ser Leu Ser Cys Arg 
    30                  35                  40 

gcc agc caa att att agc aac aac cta cac tgg tat caa caa aaa tca      195 
Ala Ser Gln Ile Ile Ser Asn Asn Leu His Trp Tyr Gln Gln Lys Ser 
45                  50                  55                  60 

cat gag tct cca agg ctt ctc atc aag tat gct tcc cag tcc atc tct      243 
His Glu Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser 
                65                  70                  75 

ggg atc ccc tcc agg ttc agt ggc agt gga tca ggg aca gat ttc act      291 
Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 
            80                  85                  90 

ctc agt atc aac agt gtg gag act gaa gat ttt gga atg tat ttc tgt      339 
Leu Ser Ile Asn Ser Val Glu Thr Glu Asp Phe Gly Met Tyr Phe Cys 
        95                  100                 105 

caa cag agt aac agc tgg cct ctc acg ttc ggc tcg ggg aca aag ctg      387 
Gln Gln Ser Asn Ser Trp Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu 
    110                 115                 120 

gag atc aaa cgg cgtaagtgtg tcagggtttc acaagaggga ctaaagacat          439 
Glu Ile Lys Arg 
125 

gtcagctaat gtgtgactaa tggtaatgtc acttgtcagg atcc                     483 

 
           
             6  
             128  
             PRT  
             Mus sp.  
           
            6 

Met Val Phe Thr Pro Gln Ile Leu Gly Leu Met Leu Phe Trp Ile Ser 
1               5                   10                  15 

Ala Ser Arg Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser 
            20                  25                  30 

Val Thr Pro Gly Asp Ser Val Ser Leu Ser Cys Arg Ala Ser Gln Ile 
        35                  40                  45 

Ile Ser Asn Asn Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro 
    50                  55                  60 

Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser 
65                  70                  75                  80 

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn 
                85                  90                  95 

Ser Val Glu Thr Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn 
            100                 105                 110 

Ser Trp Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Arg 
        115                 120                 125 

 
           
             7  
             258  
             DNA  
             Mus sp.  
             
               misc_feature  
               (1)..(258)  
               Light chain leader plus sFv of MB3.6  
             
           
            7 

gatatcagat ctcagctgtc tagacatatg gttttcacac ctcagatann nnnnnnnnnn     60 

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnngggac aaagctggag    120 

atcaaaggtg gctcaggatc gggtggagcc ggctctggtg gctcaggatc ggaagtggtg    180 

gtggtggagn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnacc    240 

acggtcaccg tctccagt                                                  258 

 
           
             8  
             682  
             DNA  
             Mus sp.  
             
               CDS  
               (20)..(418)  
               3D8 Heavy chain V region, plus leader  
             
           
            8 

tgaacacgga cccctcacc atg aac ttc ggg ctc agc ttg att ttc ctt gtc      52 
                     Met Asn Phe Gly Leu Ser Leu Ile Phe Leu Val 
                     1               5                   10 

ctt gtt tta aaa ggt gtc cag tgt gaa gtg aag gtg gtg gag tct ggg      100 
Leu Val Leu Lys Gly Val Gln Cys Glu Val Lys Val Val Glu Ser Gly 
            15                  20                  25 

gga ggc tta gtg aag cct gga gcg tct ctg aaa ctc tcc tgt gca gcc      148 
Gly Gly Leu Val Lys Pro Gly Ala Ser Leu Lys Leu Ser Cys Ala Ala 
        30                  35                  40 

tct gga ttc act ttc agt aac tat ggc atg tct tgg gtt cgc cag act      196 
Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met Ser Trp Val Arg Gln Thr 
    45                  50                  55 

tca gac aag agg ctg gag tgg gtc gca tcc att agt agt ggt ggt gat      244 
Ser Asp Lys Arg Leu Glu Trp Val Ala Ser Ile Ser Ser Gly Gly Asp 
60                  65                  70                  75 

agc acc ttc tat gca gac aat gta aag ggc cga ttc acc atc tcc aga      292 
Ser Thr Phe Tyr Ala Asp Asn Val Lys Gly Arg Phe Thr Ile Ser Arg 
                80                  85                  90 

gag aat gcc aag aac acc ctg tac ctg caa atg agt agt ctg aag tct      340 
Glu Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Lys Ser 
            95                  100                 105 

gag gac acg gcc ttg tat tac tgt gca aga gac gat cta ttt aac tgg      388 
Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Asp Asp Leu Phe Asn Trp 
        110                 115                 120 

ggc caa ggc acc act ctc aca gtc tca tca gccaaaacaa cagccccatc        438 
Gly Gln Gly Thr Thr Leu Thr Val Ser Ser 
    125                 130 

ggtctatcca ctggcccctg tgtgtggaga tacaattggc tcctcggtga ctttaggatg    498 

cctggtcaag ggttatttcc ttgagccagt gaccttgacc tggaactctg gatccctgtc    558 

cagtggtgtg cacatcttcc cagctgtctt gcagtctgac ctctacaccc tcagcagctc    618 

agtgactgta acctcgagca cctggcccag ccagtccatc acttgcaatg tggcccaccc    678 

ggca                                                                 682 

 
           
             9  
             133  
             PRT  
             Mus sp.  
           
            9 

Met Asn Phe Gly Leu Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly 
1               5                   10                  15 

Val Gln Cys Glu Val Lys Val Val Glu Ser Gly Gly Gly Leu Val Lys 
            20                  25                  30 

Pro Gly Ala Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 
        35                  40                  45 

Ser Asn Tyr Gly Met Ser Trp Val Arg Gln Thr Ser Asp Lys Arg Leu 
    50                  55                  60 

Glu Trp Val Ala Ser Ile Ser Ser Gly Gly Asp Ser Thr Phe Tyr Ala 
65                  70                  75                  80 

Asp Asn Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn 
                85                  90                  95 

Thr Leu Tyr Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Leu 
            100                 105                 110 

Tyr Tyr Cys Ala Arg Asp Asp Leu Phe Asn Trp Gly Gln Gly Thr Thr 
        115                 120                 125 

Leu Thr Val Ser Ser 
    130 

 
           
             10  
             729  
             DNA  
             Mus sp.  
             
               CDS  
               (15)..(410)  
               3D8 Light chain V region, plus leader  
             
           
            10 

ccgttgccgt cgtg atg agt cct gcc cag ttc ctg ttt ctg tta gtg ctc       50 
                Met Ser Pro Ala Gln Phe Leu Phe Leu Leu Val Leu 
                1               5                   10 

tgg att cag gaa acc aac ggt gat gtt gta atg acc cag act cca ctc       98 
Trp Ile Gln Glu Thr Asn Gly Asp Val Val Met Thr Gln Thr Pro Leu 
        15                  20                  25 

act ttg tcg gtt acc att gga caa cca gcc tct atc tct tgc aag tca      146 
Thr Leu Ser Val Thr Ile Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser 
    30                  35                  40 

agt cag agc ctc tta tat agt aat gga aaa acc tat ttg aat tgg tta      194 
Ser Gln Ser Leu Leu Tyr Ser Asn Gly Lys Thr Tyr Leu Asn Trp Leu 
45                  50                  55                  60 

tta cag agg cca ggc cag tct cca aag cgc cta atc tat ctg gtg tct      242 
Leu Gln Arg Pro Gly Gln Ser Pro Lys Arg Leu Ile Tyr Leu Val Ser 
                65                  70                  75 

aaa ctg gac tct gga gtc cct gac agg ttc act ggc agt gga tca gga      290 
Lys Leu Asp Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly 
            80                  85                  90 

aca gat ttt aca ctg aaa atc agc aga gtg gag gct gag gat ttg gga      338 
Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly 
        95                  100                 105 

gtt tat tac tgc gtg caa ggt aca cat ttt cct cac acg ttc gga ggg      386 
Val Tyr Tyr Cys Val Gln Gly Thr His Phe Pro His Thr Phe Gly Gly 
    110                 115                 120 

ggg acc aag ctg gaa ata aaa cgg gctgatgctg caccaactgt atccatcttc     440 
Gly Thr Lys Leu Glu Ile Lys Arg 
125                 130 

ccaccatcca gtgagcagtt aacatctgga ggtgcctcag tcgtgtgctt cttgaacaac    500 

ttctacccca aagacatcaa tgtcaagtgg aagattgatg gcagtgaacg acaaaatggc    560 

gtcctgaaca gttggactga tcaggacagc aaagacagca cctacagcat gagcagcacc    620 

ctcacgttga ccaaggacga gtatgaacga cataacagct atacctgtga ggccactcac    680 

aagacatcaa cttcacccat tgtcaagagc ttcaacagga atgagtgtt                729 

 
           
             11  
             132  
             PRT  
             Mus sp.  
           
            11 

Met Ser Pro Ala Gln Phe Leu Phe Leu Leu Val Leu Trp Ile Gln Glu 
1               5                   10                  15 

Thr Asn Gly Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val 
            20                  25                  30 

Thr Ile Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu 
        35                  40                  45 

Leu Tyr Ser Asn Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro 
    50                  55                  60 

Gly Gln Ser Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser 
65                  70                  75                  80 

Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr 
                85                  90                  95 

Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 
            100                 105                 110 

Val Gln Gly Thr His Phe Pro His Thr Phe Gly Gly Gly Thr Lys Leu 
        115                 120                 125 

Glu Ile Lys Arg 
    130 

 
           
             12  
             736  
             DNA  
             Mus sp.  
             
               CDS  
               (14)..(430)  
               4D4 Heavy chain V region, plus leader  
             
           
            12 

actgactcta acc atg gga tgg aga tgg atc ttt ctt ttc ctc ctg tca        49 
               Met Gly Trp Arg Trp Ile Phe Leu Phe Leu Leu Ser 
               1               5                   10 

gga act gca ggt gtc cat tgc cag gtt cag ctg cag cag tct gga cct       97 
Gly Thr Ala Gly Val His Cys Gln Val Gln Leu Gln Gln Ser Gly Pro 
        15                  20                  25 

gag ctg gtg aag cct ggg gct tta gtg aag ata tcc tgc aag gct tct      145 
Glu Leu Val Lys Pro Gly Ala Leu Val Lys Ile Ser Cys Lys Ala Ser 
    30                  35                  40 

ggt tac acc ttc aca agc tac gat ata aac tgg gtg aag cag agg cct      193 
Gly Tyr Thr Phe Thr Ser Tyr Asp Ile Asn Trp Val Lys Gln Arg Pro 
45                  50                  55                  60 

gga cag gga ctt gag tgg att gga tgg att tat cct gga gat ggt ggt      241 
Gly Gln Gly Leu Glu Trp Ile Gly Trp Ile Tyr Pro Gly Asp Gly Gly 
                65                  70                  75 

act aat tac aat gag aaa ttc aag ggc aag gcc aca ctg act gca gac      289 
Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp 
            80                  85                  90 

aaa tcc tcc agc aca gcc tac atg cag ctc agt agc ctg act tct gag      337 
Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu 
        95                  100                 105 

aac tct gca gtc tat ttc tgt gca aga ggg ggt aac ttc cct tct tat      385 
Asn Ser Ala Val Tyr Phe Cys Ala Arg Gly Gly Asn Phe Pro Ser Tyr 
    110                 115                 120 

gct atg gac tac tgg ggt caa gga acc tca gtc acc gtc tcc tca          430 
Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 
125                 130                 135 

gccaaaacga cacccccatc tgtctatcca ctggcccctg gatctgctgc ccaaactaac    490 

tccatggtga ccccgggatg cctggtcaag ggctatttcc ctgagccagt gacagtgacc    550 

tggaactctg gatccctgtc cagcggtgtg cacaccttcc cagctgtcct gcagtctgac    610 

ctctacactc tgagcagctc agtgactgtc ccctccagca cctggcccag cgagaccgtc    670 

acctgcaacg ttgcccaccc ggccagcagc accaaggtgg acaagaaaat tgtgcccagg    730 

gattgt                                                               736 

 
           
             13  
             139  
             PRT  
             Mus sp.  
           
            13 

Met Gly Trp Arg Trp Ile Phe Leu Phe Leu Leu Ser Gly Thr Ala Gly 
1               5                   10                  15 

Val His Cys Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys 
            20                  25                  30 

Pro Gly Ala Leu Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe 
        35                  40                  45 

Thr Ser Tyr Asp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 
    50                  55                  60 

Glu Trp Ile Gly Trp Ile Tyr Pro Gly Asp Gly Gly Thr Asn Tyr Asn 
65                  70                  75                  80 

Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser 
                85                  90                  95 

Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asn Ser Ala Val 
            100                 105                 110 

Tyr Phe Cys Ala Arg Gly Gly Asn Phe Pro Ser Tyr Ala Met Asp Tyr 
        115                 120                 125 

Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 
    130                 135 

 
           
             14  
             504  
             DNA  
             Mus sp.  
             
               CDS  
               (7)..(402)  
               4D4 Light chain V region, plus leader  
             
           
            14 

ctcaaa atg aag ttg cct gtt agg ctg ttg gtg ctg atg ttc tgg att        48 
       Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile 
       1               5                   10 

cct gct tcc aac agt gat gtt ttg atg acc caa tct cca ctc tcc ctg       96 
Pro Ala Ser Asn Ser Asp Val Leu Met Thr Gln Ser Pro Leu Ser Leu 
15                  20                  25                  30 

cct gtc agt ctt gga gat caa gcc tcc atc tct tgc aga tct agt cag      144 
Pro Val Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln 
                35                  40                  45 

agc att gtc cat agt aat gga gac acc tat tta gaa tgg tac ctg cag      192 
Ser Ile Val His Ser Asn Gly Asp Thr Tyr Leu Glu Trp Tyr Leu Gln 
            50                  55                  60 

aaa cca ggc cag tct cca aag ctc ctg atc tac aag gtt tcc gac cga      240 
Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asp Arg 
        65                  70                  75 

ttt tct ggg gtc cca gac agg ttc agt ggc agt gga tca ggg aca gat      288 
Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 
    80                  85                  90 

ttc aca ctc aag atc agc aga gtg gag gct gag gat ctg gga gtt tat      336 
Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr 
95                  100                 105                 110 

ttc tgc ttt caa ggt tca cat gtt ccg tac gcg ttc gga ggg ggg acc      384 
Phe Cys Phe Gln Gly Ser His Val Pro Tyr Ala Phe Gly Gly Gly Thr 
                115                 120                 125 

aag ctg gaa ata aaa cgg gctgatgctg caccaactgt atccatcttc             432 
Lys Leu Glu Ile Lys Arg 
            130 

ccaccatcca gtgagcagtt aacatctgga ggtgcctcag tcgtgtgctt cttgaacaac    492 

ttctacccca aa                                                        504 

 
           
             15  
             132  
             PRT  
             Mus sp.  
           
            15 

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 
1               5                   10                  15 

Ser Asn Ser Asp Val Leu Met Thr Gln Ser Pro Leu Ser Leu Pro Val 
            20                  25                  30 

Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 
        35                  40                  45 

Val His Ser Asn Gly Asp Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 
    50                  55                  60 

Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asp Arg Phe Ser 
65                  70                  75                  80 

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 
                85                  90                  95 

Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys 
            100                 105                 110 

Phe Gln Gly Ser His Val Pro Tyr Ala Phe Gly Gly Gly Thr Lys Leu 
        115                 120                 125 

Glu Ile Lys Arg 
    130 

 
           
             16  
             761  
             DNA  
             Mus sp.  
             
               CDS  
               (62)..(478)  
               3E11 Heavy chain V region, plus leader  
             
           
            16 

cctggattca atttccagtt cctcacattc agtgatcagc actgaacacg gacccctcac     60 

c atg aac ttc ggg ctc agc ttg att ttc ctt gtc ctt gtt tta aaa ggt    109 
  Met Asn Phe Gly Leu Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly 
  1               5                   10                  15 

gtc cag tgt gaa gtg aaa ctg gtg gag tct ggg gga gac tta atg aac      157 
Val Gln Cys Glu Val Lys Leu Val Glu Ser Gly Gly Asp Leu Met Asn 
            20                  25                  30 

cct gga gcg tct ctg aaa ctc tcc tgt gca gcc tct gga ttc agt ttc      205 
Pro Gly Ala Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe 
        35                  40                  45 

agt aac tat ggc atg tct tgg gtt cgc cag act tca gac aag agg ctg      253 
Ser Asn Tyr Gly Met Ser Trp Val Arg Gln Thr Ser Asp Lys Arg Leu 
    50                  55                  60 

gag tgg gtc gct tcc att agt acg ggt ggt gct aat acc ttc tat cca      301 
Glu Trp Val Ala Ser Ile Ser Thr Gly Gly Ala Asn Thr Phe Tyr Pro 
65                  70                  75                  80 

gac aat gta aag ggc cga ttc acc att tcc aga gag aat gcc aag aac      349 
Asp Asn Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn 
                85                  90                  95 

acc cta tac ctg caa atg agt agt ctg aag tct gag gac acg gcc ttg      397 
Thr Leu Tyr Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Leu 
            100                 105                 110 

tat ttc tgt gca aga gat agt cac tcc gta ggt tgt tgg ttt gct acc      445 
Tyr Phe Cys Ala Arg Asp Ser His Ser Val Gly Cys Trp Phe Ala Thr 
        115                 120                 125 

tgg ggc caa ggg act ctg gtc act gtc tct gca gccaaaacaa cacccccatc    498 
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 
    130                 135 

agtctatcca ctggcccctg ggtgtggaga tactactggt tcctccgtga ctctgggatg    558 

cctggtcaag ggctacttcc ctgagtcagt gactgtgact tggaactccg gatccctgcc    618 

cagcagtgtg cacaccttcc cagctctcct gcagtctgga ctctacacta tgagcagctc    678 

agtgactgtc ccctccagca cctggccaag ccagaccgtt acctgcagtg ttgctcaccc    738 

agccagcagc accacggtgg aca                                            761 

 
           
             17  
             139  
             PRT  
             Mus sp.  
           
            17 

Met Asn Phe Gly Leu Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly 
1               5                   10                  15 

Val Gln Cys Glu Val Lys Leu Val Glu Ser Gly Gly Asp Leu Met Asn 
            20                  25                  30 

Pro Gly Ala Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe 
        35                  40                  45 

Ser Asn Tyr Gly Met Ser Trp Val Arg Gln Thr Ser Asp Lys Arg Leu 
    50                  55                  60 

Glu Trp Val Ala Ser Ile Ser Thr Gly Gly Ala Asn Thr Phe Tyr Pro 
65                  70                  75                  80 

Asp Asn Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn 
                85                  90                  95 

Thr Leu Tyr Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Leu 
            100                 105                 110 

Tyr Phe Cys Ala Arg Asp Ser His Ser Val Gly Cys Trp Phe Ala Thr 
        115                 120                 125 

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 
    130                 135 

 
           
             18  
             698  
             DNA  
             Mus sp.  
             
               CDS  
               (6)..(401)  
               3E11 Light chain V region, plus leader  
             
           
            18 

ccagc atg ggc atc aag atg gaa tca cag act ctg gtc ttc ata tcc ata     50 
      Met Gly Ile Lys Met Glu Ser Gln Thr Leu Val Phe Ile Ser Ile 
      1               5                   10                  15 

ctg ctc tgg tta tat gga gct gat ggg aac att gta atg acc caa tct       98 
Leu Leu Trp Leu Tyr Gly Ala Asp Gly Asn Ile Val Met Thr Gln Ser 
                20                  25                  30 

ccc aaa tcc atg tcc atg tca gta gga gag agg gtc acc ttg acc tgc      146 
Pro Lys Ser Met Ser Met Ser Val Gly Glu Arg Val Thr Leu Thr Cys 
            35                  40                  45 

aag gcc agt gag aat gtg gtt act tat gtt tcc tgg tat caa cag aaa      194 
Lys Ala Ser Glu Asn Val Val Thr Tyr Val Ser Trp Tyr Gln Gln Lys 
        50                  55                  60 

cca gag cag tct cct aaa ctg ctg ata tac ggg gca tcc aac cgg tac      242 
Pro Glu Gln Ser Pro Lys Leu Leu Ile Tyr Gly Ala Ser Asn Arg Tyr 
    65                  70                  75 

act ggg gtc ccc gat cgc ttc aca ggc agt gga tct gca aca gat ttc      290 
Thr Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Ala Thr Asp Phe 
80                  85                  90                  95 

act ctg acc atc agc agt gtg cag gct gaa gac ctt gca gat tat cac      338 
Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr His 
                100                 105                 110 

tgt gga cag ggt tac agc tat ccg tac acg ttc gga ggg ggg acc aag      386 
Cys Gly Gln Gly Tyr Ser Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 
            115                 120                 125 

ctg gaa ata aaa cgg gctgatgctg caccaactgt atccatcttc ccaccatcca      441 
Leu Glu Ile Lys Arg 
        130 

gtgagcagtt aacatctgga ggtgcctcag tcgtgtgctt cttgaacaac ttctacccca    501 

aagacatcaa tgtcaagtgg aagattgatg gcagtgaacg acaaaatggc gtcctgaaca    561 

gttggactga tcaggacagc aaagacagca cctacagcat gagcagcacc ctcacgttga    621 

ccaaggacga gtatgaacga cataacagct atacctgtga ggccactcac aagacatcaa    681 

cttcacccat cgtcaag                                                   698 

 
           
             19  
             132  
             PRT  
             Mus sp.  
           
            19 

Met Gly Ile Lys Met Glu Ser Gln Thr Leu Val Phe Ile Ser Ile Leu 
1               5                   10                  15 

Leu Trp Leu Tyr Gly Ala Asp Gly Asn Ile Val Met Thr Gln Ser Pro 
            20                  25                  30 

Lys Ser Met Ser Met Ser Val Gly Glu Arg Val Thr Leu Thr Cys Lys 
        35                  40                  45 

Ala Ser Glu Asn Val Val Thr Tyr Val Ser Trp Tyr Gln Gln Lys Pro 
    50                  55                  60 

Glu Gln Ser Pro Lys Leu Leu Ile Tyr Gly Ala Ser Asn Arg Tyr Thr 
65                  70                  75                  80 

Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Ala Thr Asp Phe Thr 
                85                  90                  95 

Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr His Cys 
            100                 105                 110 

Gly Gln Gly Tyr Ser Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu 
        115                 120                 125 

Glu Ile Lys Arg 
    130