Patent Publication Number: US-2022227871-A1

Title: Anti-pd-l1 antibody

Description:
PRIORITY APPLICATIONS 
     This application is a continuation of U.S. application Ser. No. 16/325,040, Filed Feb. 12, 2019, which is a U.S. National Stage Filing under 35 U.S.C. 371 from International Application No. PCT/JP2017/029055, filed on Aug. 10, 2017, and published as WO2018/034225 on Feb. 22, 2018, which claims the benefit of priority to Japanese Application No. 2017-110723, filed on Jun. 5, 2017 and to Japanese Application No. 2017-061454, filed on Mar. 27, 2017 and to Japanese Application No. 2016-159089, filed on Aug. 15, 2016 and to Japanese Application No. 2016-159088, filed on Aug. 15, 2016; the benefit of priority of each of which is hereby claimed herein, and which applications and publication are hereby incorporated herein by reference in their entireties. 
    
    
     TECHNICAL FIELD 
     The present invention relates to an anti-PD-L1 antibody. More specifically, the present invention relates to an anti-PD-L1 antibody comprising a variable region containing complementarity-determining regions (CDR) of a rat anti-bovine PD-L1 antibody and a constant region of an antibody of an animal other than rat. 
     BACKGROUND ART 
     Programmed cell death 1 (PD-1), an immunoinhibitory receptor, and its ligand programmed cell death ligand 1 (PD-L1) are molecules identified by Prof. Tasuku Honjo et al., Kyoto University, as factors which inhibit excessive immune response and are deeply involved in immunotolerance (Non-Patent Document No. 1: Ishida Y, Agata Y, Shibahara K, Honjo T The EMBO Journal. 1992 November; 11(11):3887-3895). Recently, it has been elucidated that these molecules are also involved in immunosuppression in tumors. In the field of human medical care, an antibody drug that inhibits the effect of PD-1 has been developed and put into practical use (Opdivo™, Ono Pharmaceutical Co., Ltd.) 
     To date, the present inventors have been developing an immunotherapy for animal refractory diseases targeting PD-1 or PD-L1, and have revealed that this novel immunotherapy is applicable to multiple-diseases and multiple-animals. (Non-Patent Document No. 2: Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y, Murata S, Ohashi K. Immunology. 2014 August; 142(4):551-61: Non-Patent Document No. 3: Maekawa N. Konnai S, Ikebuchi R, Okagawa T. Adachi M, Takagi S. Kagawa Y, Nakajima C. Suzuki Y, Murata S, Ohashi K. PLoS One. 2014 Jun. 10; 9(6):e98415; Non-Patent Document No. 4: Mingala C N, Konnai S, Ikebuchi R. Ohashi K. Comp. Immunol. Microbiol. Infect. Dis. 2011 January; 34(1):55-63.) 
     However, the antibodies which the present inventors have prepared to date are rat antibodies, and therefore it is impossible to administer those antibodies repeatedly to animals other than rat. 
     PRIOR ART LITERATURE 
     Non-Patent Documents 
     
         
         Non-Patent Document No. 1: Ishida Y. Agata Y, Shibahara K, Honjo T The EMBO Journal. 1992 November; 11(11):3887-3895. 
         Non-Patent Document No. 2: Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y. Murata S, Ohashi K. Immunology. 2014 August; 142(4):551-61. 
         Non-Patent Document No. 3: Maekawa N, Konnai S, Ikebuchi R. Okagawa T, Adachi M, Takagi S. Kagawa Y. Nakajima C, Suzuki Y, Murata S. Ohashi K. PLoS One. 2014 Jun. 10; 9(6):e98415. 
         Non-Patent Document No. 4: Mingala C N, Konnai S, Ikebuchi R, Ohashi K. Comp. Immunol. Microbiol. Infect. Dis. 2011 January; 34(1):55-63. 
       
    
     DISCLOSURE OF THE INVENTION 
     Problem for Solution by the Invention 
     It is an object of the present invention to provide an anti-PD-L1 antibody capable of repeated administration even to animals other than rat. 
     Means to Solve the Problem 
     The present inventors have determined the variable regions of a rat anti-bovine PD-L1 monoclonal antibody (4G12) capable of inhibiting the binding of canine PD-1 to PD-L1, and then combined genes encoding the resultant variable regions with genes encoding the constant regions of a canine immunoglobulin (IgG-D equivalent to human IgG4) to thereby obtain a chimeric antibody gene, which was introduced into Chinese hamster ovary cells (CHO cells). By culturing/proliferating the resultant CHO cells, the present inventors have succeeded in preparing a rat-canine chimeric anti-PD-L1 antibody. Further, the present inventors have determined the CDRs of the variable region of the rat anti-bovine PD-L1 monoclonal antibody 4G12. 
     Furthermore, the present inventors have determined the variable regions of the rat anti-bovine PD-L1 monoclonal antibody 4G12 capable of inhibiting the binding of bovine PD-1 to PD-L1, and then combined genes encoding the resultant variable regions with genes encoding the constant regions of a bovine immunoglobulin (bovine IgG1, with mutations having been introduced into the putative binding sites of Fcγ receptors in CH2 domain in order to inhibit ADCC activity; see  FIG. 19  for amino acid numbers and mutations: 250 E→P, 251 L→V, 252 P→A, 253 G→deletion, 347 A→S, 348 P→S; Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y, Murata S, Ohashi K. Immunology 2014 August; 142(4):551-561) to thereby obtain a chimeric antibody gene. This gene was introduced into Chinese hamster ovary cells (CHO cells). By culturing/proliferating the resultant cells, the present inventors have succeeded in preparing a rat-bovine chimeric anti-PD-L1 antibody. The present invention has been achieved based on these findings. 
     A summary of the present invention is as described below.
     (1) An anti-PD-L1 antibody comprising (a) a light chain comprising a light chain variable region containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the light chain constant region of an antibody of an animal other than rat; and (b) a heavy chain comprising a heavy chain variable region containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the heavy chain constant region of an antibody of an animal other than rat.   (2) The antibody of (1) above, wherein the light chain variable region and the heavy chain variable region are derived from rat.   (3) The antibody of (2) above, wherein the light chain variable region is the light chain variable region of a rat anti-bovine PD-L1 antibody and the heavy chain variable region is the heavy chain variable region of a rat anti-bovine PD-L1 antibody.   (4) The antibody of (3) above, wherein the light chain variable region has the amino acid sequence as shown in SEQ ID NO. 1 and the heavy chain variable region has the amino acid sequence as shown in SEQ ID NO: 2.   (5) The antibody of any one of (1) to (4) above, wherein the light chain constant region of an antibody of an animal other than rat has the amino acid sequence of the constant region of lambda chain or kappa chain.   (6) The antibody of any one of (1) to (5) above, wherein the heavy chain constant region of an antibody of an animal other than rat has the amino acid sequence of the constant region of an immunoglobulin equivalent to human IgG4.   (7) The antibody of any one of (1) to (5) above, wherein the animal other than rat is bovine and the heavy chain constant region of the bovine antibody has mutations introduced thereinto that reduce ADCC activity and/or CDC activity.   (8) The antibody of (6) above, wherein the animal other than rat is canine; the light chain constant region of the canine antibody has the amino acid sequence of the constant region of lambda chain; and the heavy chain constant region of the canine antibody has the amino acid sequence of the constant region of an immunoglobulin equivalent to human IgG4.   (9) The antibody of (7) above, wherein the light chain constant region of the bovine antibody has the amino acid sequence of the constant region of lambda chain and the heavy chain constant region of the bovine antibody has mutations introduced thereinto that reduce ADCC activity and/or CDC activity.   (10) The antibody of (8) above, wherein the light chain constant region of the canine antibody has the amino acid sequence as shown in SEQ ID NO: 3 and the heavy chain constant region of the canine antibody has the amino acid sequence as shown in SEQ ID NO: 4.   (11) The antibody of (9) above, wherein the light chain constant region of the bovine antibody has the amino acid sequence as shown in SEQ ID NO: 100 and the heavy chain constant region of the bovine antibody has the amino acid sequence as shown in SEQ ID NO: 102.   (12) The antibody of any one of (1) to (11) above which has a four-chain structure comprising two light chains and two heavy chains.   (13) A pharmaceutical composition comprising the antibody of any one of (1) to (12) above as an active ingredient.   (14) The composition of (13) above for prevention and/or treatment of cancers and/or inflammations.   (15) The composition of (14) above, wherein the cancers and/or inflammations are selected from the group consisting of neoplastic diseases, leukemia, Johne&#39;s disease, anaplasmosis, bacterial mastitis, mycotic mastitis, mycoplasma infections (such as mycoplasma mastitis, mycoplasma pneumonia or the like), tuberculosis,  Theileria orientalis  infection, cryptosporidiosis, coccidiosis, trypanosomiasis and leishmaniasis.   (16) An artificial genetic DNA comprising (a′) a DNA encoding a light chain comprising a light chain variable region containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the light chain constant region of an antibody of an animal other than rat and (b′) a DNA encoding a heavy chain comprising a heavy chain variable region containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the heavy chain constant region of an antibody of an animal other than rat.   (17) A vector comprising the artificial genetic DNA of (16) above.   (18) A host cell transformed with the vector of (17) above.   (19) A method of preparing an antibody, comprising culturing the host cell of (18) above and collecting an anti-PD-L1 antibody from the resultant culture.   (20) A DNA encoding a light chain comprising a light chain variable region containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the light chain constant region of an antibody of an animal other than rat.   (21) A DNA encoding a heavy chain comprising a heavy chain variable region containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the heavy chain constant region of an antibody of an animal other than rat.   

     The present specification encompasses the contents disclosed in the specifications and/or drawings of Japanese Patent Applications No. 2016-159088, No. 2016-159089, No. 2017-110723 and No. 2017-61454 based on which the present patent application claims priority. 
     Effect of the Invention 
     According to the present invention, a novel anti-PD-L1 antibody has been obtained. This antibody is applicable even to those animals other than rat. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. 
         FIG. 1  Inhibition of the binding of recombinant canine PD-L1 to recombinant canine PD-1. The binding of canine PD-L1-Ig to canine PD-1-Ig was detected on ELISA plates. The optical density (O.D.) without addition of antibody was taken as 100%. O.D. at each antibody concentration was shown as relative value. Among rat anti-bovine PD-L1 monoclonal antibodies 4G12 (Rat IgG2a (κ)), 5A2 (Rat IgG1 (κ)) and 6G7 (Rat IgM (κ)) which showed cross-reaction with canine PD-L1, clones 4G12 and 6G7 exhibited a high binding inhibition capacity. 
         FIG. 2  Schematic drawings of pDC6 vector and a rat-canine chimeric anti-PD-L1 antibody. 
         FIG. 3  Expression and purification of rat-canine chimeric anti-PD-L1 antibodies c4G12 and c6G7. SDS-PAGE was performed under non-reducing conditions, followed by visualization of bands by CBB staining. a: purification with protein A alone. b: a +gel filtration chromatography. 
         FIG. 4  PD-1/PD-L1 binding inhibition activities of rat-canine chimeric anti-PD-L1 antibodies c4G12 and c6G7. 
         FIG. 5  Establishment of cell clones capable of high expression of rat-canine chimeric anti-PD-L1 antibody c4G12. 
         FIG. 6  SDS-PAGE images of rat-canine chimeric anti-PD-L1 antibody c4G12. Rat anti-bovine PD-L1 antibody 4G12 and rat-canine chimeric anti-PD-L1 antibody c4G12 were electrophoresed under reducing conditions and non-reducing conditions, followed by visualization of bands by CBB staining. Under reducing conditions, a band of antibody&#39;s heavy chain was detected at around 50 kDa and a band of antibody&#39;s light chain at around 25 kDa. No bands other than the bands of interest were detected. 
         FIG. 7  Inhibitory activities of rat anti-bovine PD-L1 antibody 4G12 and rat-canine chimeric anti-PD-L1 antibody c4G12 against canine PD-1/PD-L1 binding and CD80/PD-L1 binding. Rat anti-bovine PD-L1 monoclonal antibody 4G12 and rat-canine chimeric anti-PD-L1 antibody c4G12 reduced the amounts of binding of PD-L1-Ig to canine PD-1-Ig and CD80-Ig. No change due to chimerization of the antibody was observed in binding inhibition activity 
         FIG. 8  Canine immune cell activation effect by rat-canine chimeric anti-PD-L1 antibody c4G12. Canine PBMCs were cultured under stimulation for 3 days, followed by determination of IL-2 and IFN-γ concentrations in the supernatant by ELISA. Further, nucleic acid analogue EdU was added to the culture medium at day 2 of the culture under stimulation, followed by determination of the EdU uptake by flow cytometry. Rat-canine chimeric anti-PD-L1 antibody c4G12 increased the production of IL-2 and IFN-γ from canine PBMCs and enhanced proliferation of CD4+ and CD8 +  lymphocytes. 
         FIG. 9  Expression of PD-L1 in oral melanoma (A) and undifferentiated sarcoma (B) 
         FIG. 10  CT images and appearances of tumor in a test of treatment by administering rat-canine chimeric anti-PD-L1 antibody c4G12 to a dog with oral melanoma. (a,d) Before the start of the treatment, (b,e) at week 10 of the treatment, and (cf) at week 34 of the treatment. A remarkable anti-tumor effect was recognized upon five administrations of the antibody (at week 10 from the start of the treatment). At week 34, a further reduction of tumor was confirmed. 
         FIG. 11  Time-dependent changes in the longest diameter of the tumor in the dog with oral melanoma shown in  FIG. 10 . Reduction by 30% or more compared to the baseline longest diameter was regarded as partial response (PR). 
         FIG. 12  CT images in a test of treatment by administering rat-canine chimeric anti-PD-L1 antibody c4G12 to a dog with undifferentiated sarcoma. (a,c) Before the start of the treatment, (b,d) at week 3 of the treatment. A remarkable reduction of tumor was recognized upon two administrations of the antibody. 
         FIG. 13  CT images in a test of treatment by administering rat-canine chimeric anti-PD-L1 antibody c4G12 to dogs with oral melanoma (pulmonary metastatic cases). (a,d,g) Before the start of the treatment, (b,e,h) at week 6 of the treatment, and (c,f,i) at week 18 of the treatment. A plurality of pulmonary metastatic lesions disappeared upon nine administrations of the antibody. 
         FIG. 14  Time-dependent changes in the proportion survival of dogs with oral melanoma after the occurrence of pulmonary metastasis. In the antibody administration group, the survival duration may have been prolonged compared to the control group. 
         FIG. 15  CDR1, CDR2 and CDR3 regions in the light chain variable region and the heavy chain variable region of rat anti-bovine PD-L1 antibody 4G12 are illustrated. 
         FIG. 16  Cross-reactivity of rat anti-bovine PD-L1 antibody 4012. It was confirmed that rat anti-bovine PD-L1 antibody 4G12 binds to ovine PD-L1 and porcine PD-L1. 
         FIG. 17  Reactivity of rat anti-bovine PD-L1 antibody 4G12 with water buffalo leukocytes. Rat anti-bovine PD-L1 antibody 4G12 strongly bound to blood macrophages (CD14 +  CD11b +  cells) of water buffalo, whereas rat anti-bovine PD-L1 antibody 4G12 bound weakly to lymphocytes (CD14 − CD11b −  cells) of water buffalo. It is believed that this difference in binding reflects the expression levels of PD-L1 in macrophages and lymphocytes. 
         FIG. 18  Inhibition test on rat anti-bovine PD-L1 antibody 4G12 against ovine or porcine PD-1/PD-L1 binding. It was demonstrated that rat anti-bovine PD-L1 antibody 4G12 is capable of inhibiting ovine and porcine PD-1/PD-L1 binding in a concentration dependent manner. 
         FIG. 19  The amino acid sequence of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. CDR1, CDR2 and CDR3 regions in the light chain variable region and the heavy chain variable region of rat anti-bovine PD-L1 antibody 4G12 are shown. Further, amino acids introduced as mutations to bovine IgG1 (CH2 domain) are also shown (amino acid numbers and mutations: 250 E→P, 251 L→V, 252 P→A, 253 G→deletion, 347 A→S, 348 P→S). 
         FIG. 20  Schematic drawings of pDC6 vector and rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. 
         FIG. 21  Confirmation of the purity of purified rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. 
         FIG. 22  Binding specificity of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. 
         FIG. 23  Inhibitory activity of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 against bovine PD-1/PD-L1 binding (the test results of inhibition against binding of bovine PD-L1 expressing cells and soluble bovine PD-1). 
         FIG. 24  Inhibitory activity of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 against bovine PD-1/PD-L1 binding (the test results of inhibition against binding of bovine PD-1 expressing cells and soluble bovine PD-L1). 
         FIG. 25  Activation effect of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 on bovine lymphocyte response (in terms of cell proliferation). 
         FIG. 26  Activation effect of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 on bovine lymphocyte response to BLV antigen (in terms of IFN-γ production). 
         FIG. 27  The proliferation response of T cells against BLV antigen in a calf experimentally infected with BLV through administration of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. 
         FIG. 28  Changes in BLV provirus loads in the calf experimentally infected with BLV through administration of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. 
     
    
    
     BEST MODES FOR CARRYING OUT THE INVENTION 
     Hereinbelow, the present invention will be described in detail. 
     The present invention provides an anti-PD-L1 antibody comprising (a) a light chain comprising a light chain variable region containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and a light chain constant region of an antibody of an animal other than rat; and (b) a heavy chain comprising a heavy chain variable region containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and a heavy chain constant region of an antibody of an animal other than rat. 
     CDR1, CDR2 and CDR3 in the light chain variable region (VL) of rat anti-bovine PD-L1 antibody 4G12 are a region consisting of the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), a region consisting of the amino acid sequence of WAT and a region consisting of the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38), respectively (see  FIG. 15 ). 
     Further, CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) of rat anti-bovine PD-L1 antibody 4G12 are a region consisting of the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), a region consisting of the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and a region consisting of the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41), respectively (see  FIG. 15 ). 
     In the amino acid sequences of QSLLYSENQKDY (SEQ ID NO: 37), WAT and GQYLVYPFT (SEQ ID NO: 38), as well as the amino acid sequences of GYTFTSNF (SEQ ID NO: 39), IYPEYGNT (SEQ ID NO: 40) and ASEEAVISLVY (SEQ ID NO: 41), one, two, three, four or five amino acids may be deleted, substituted or added. 
     As used herein, the term “antibody” is a concept encompassing not only full-length antibodies but also antibodies of smaller molecular sizes such as Fab, F(ab)′ 2 , ScFv, Diabody, V H , V L , Sc(Fv) 2 , Bispecific sc(Fv) 2 , Minibody, scFv-Fc monomer and scFv-Fc dimer. 
     In the anti-PD-L1 antibody of the present invention, VL and VH thereof may be derived from rat. For example, VL thereof may be the VL of a rat anti-bovine PD-L1 antibody, and VH thereof may be the VH of the rat anti-bovine PD-L1 antibody. 
     The amino acid sequence of the VL and the amino acid sequence of the VH of the rat anti-bovine PD-L1 antibody are shown in SEQ ID NOS: 1 and 2, respectively. The amino acid sequences as shown in SEQ ID NOS: 1 and 2 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) amino acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as VL or VH of the PD-L1 antibody. 
     The VL and VH of an antibody of an animal other than rat may be derived from an animal which produces a PD-L1 that cross-reacts with rat anti-bovine PD-L1 antibody 4G12. 
     There are two types of immunoglobulin light chain, which are called Kappa chain (κ) and Lambda chain (λ). In the anti-PD-L1 antibody of the present invention, the light chain constant region (CL) of an antibody of an animal other than rat may have the amino acid sequence of the constant region of either Kappa chain or Lambda chain. However, the relative abundance of Lambda chain is higher in ovine, feline, canine, equine and bovine, and that of Kappa chain is higher in mouse, rat, human and porcine. Since a chain with a higher relative abundance is considered to be preferable, an ovine, feline, canine, equine or bovine antibody preferably has the amino acid sequence of the constant region of Lambda chain whereas a mouse, rat, human or porcine antibody preferably has the amino acid sequence of the constant region of Kappa chain. 
     The heavy chain constant region (CH) of an antibody of an animal other than rat may have the amino acid sequence of the constant region of an immunoglobulin equivalent to human IgG4. Immunoglobulin heavy chain is classified into γ chain, μ chain, α chain, δ chain and ε chain depending on the difference in constant region. According to the type of heavy chain present, five classes (isotypes) of immunoglobulin are formed, they are IgG, IgM, IgA, IgD and IgE. 
     Immunoglobulin G (IgG) accounts for 70-75% of human immunoglobulins and is the most abundantly found monomeric antibody in plasma IgG has a four-chain structure consisting of two light chains and two heavy chains. Human IgG1, IgG2 and IgG4 have molecular weights of about 146,000, whereas human IgG3 has a long hinge region that connects Fab region and Fc region and has a larger molecular weight of 170,000. Human IgG1 accounts for about 65%, human IgG2 about 25%, human IgG3 about 7%, and human IgG4 about 3% of human IgG. They are uniformly distributed inside and outside of blood vessels. Having a strong affinity for Fc receptors and complement factors on effector cell surfaces, human IgG1 induces antibody-dependent cell cytotoxicity (ADCC) and also activates complements to induce complement-dependent cell cytotoxicity (CDC). Human IgG2 and IgG4 are low at ADCC and CDC activities because their affinity for Fc receptors and complement factors is low. 
     Immunoglobulin M (IgM), which accounts for about 10% of human immunoglobulins, is a pentameric antibody consisting of five basic four-chain structures joined together. It has a molecular weight of 970,000. Usually occurring only in blood, IgM is produced against infectious microorganisms and takes charge of early stage immunity. 
     Immunoglobulin A (IgA) accounts for 10-15% of human immunoglobulins. It has a molecular weight of 160,000. Secreted IgA is a dimeric antibody consisting of two IgA molecules joined together. IgA1 is found in serum, nasal discharge, saliva and breast milk. In intestinal juice, IgA2 is found abundantly. 
     Immunoglobulin D (IgD) is a monomeric antibody accounting for no more than 1% of human immunoglobulins. IgD is found on B cell surfaces and involved in induction of antibody production. 
     Immunoglobulin E (IgE) is a monomeric antibody that occurs in an extremely small amount, accounting for only 0.001% or less of human immunoglobulins. Immunoglobulin E is considered to be involved in immune response to parasites but in advanced countries where parasites are rare. IgE is largely involved in bronchial asthma and allergy among other things. 
     With respect to canine, sequences of IgG-A (equivalent to human IgG2), IgG-B (equivalent to human IgG1), IgG-C (equivalent to human IgG3) and IgG-D (equivalent to human IgG4) have been identified as the heavy chain of IgG. In the antibody of the present invention, an IgG&#39;s heavy chain constant region with neither ADCC activity nor CDC activity is preferable (IgG4 in human). In the case where the constant region of an immunoglobulin equivalent to human IgG4 has not been identified, one may use a constant region that has lost both ADCC activity and CDC activity as a result of introducing mutations into the relevant region of an immunoglobulin equivalent to human IgG4. 
     With respect to bovine, sequences of IgG1, IgG2 and IgG3 have been identified as the heavy chain of IgG. In the antibody of the present invention, an IgG&#39;s heavy chain constant region with neither ADCC activity nor CDC activity is preferable (IgG4 in human). Although the constant region of wild-type human IgG1 has ADCC activity and CDC activity, it is known that these activities can be reduced by introducing amino acid substitutions or deletions into specific sites. In bovine, the constant region of an immunoglobulin equivalent to human IgG4 has not been identified, so mutations may be added at the relevant region of an immunoglobulin equivalent to human IgG1 and the resultant constant region then used. As one example, the amino acid sequence of the CH of a bovine antibody (IgG1 chain, GenBank: X62916) having mutations introduced into CH2 domain and a nucleotide sequence for such amino acid sequence (after codon optimization) are shown in SEQ ID NOS: 102 and 102, respectively. 
     When an animal other than rat is canine, an anti-PD-L1 antibody is more preferable in which (i) the CL of a canine antibody has the amino acid sequence of the constant region of Lambda chain and (ii) the CH of the canine antibody has the amino acid sequence of the constant region of an immunoglobulin equivalent to human IgG4. 
     When an animal other than rat is bovine, an anti-PD-L1 antibody is more preferable in which (i) the CL of a bovine antibody has the amino acid sequence of the constant region of Lambda chain and (ii) the CH of the bovine antibody has mutations introduced thereinto that reduce ADCC activity and/or CDC activity. 
     The anti-PD-L1 antibody of the present invention encompasses rat-canine chimeric antibodies, caninized antibodies, complete canine-type antibodies, rat-bovine chimeric antibodies, bovinized antibodies and complete bovine-type antibodies. However, animals are not limited to canine and bovine and may be exemplified by human, porcine, simian, mouse, feline, equine, goat, sheep, water buffalo, rabbit, hamster, guinea pig and the like. 
     For example, the anti-PD-L1 antibody of the present invention may be an anti-PD-L1 antibody in which the CL of a canine antibody has the amino acid sequence as shown in SEQ ID NO: 3 and the CH of the canine antibody has the amino acid sequence as shown in SEQ ID NO: 4. 
     As a further example, the anti-PD-L1 antibody of the present invention may be an anti-PD-L1 antibody in which the CL of a bovine antibody has the amino acid sequence as shown in SEQ ID NO: 100 and the CH of the bovine antibody has the amino acid sequence as shown in SEQ ID NO: 102. 
     The amino acid sequences as shown in SEQ ID NOS: 3 and 4 as well as SEQ ID NOS: 100 and 102 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) amino acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as CL or CH of the PD-L1 antibody. 
     The anti-PD-L1 antibody of the present invention may have a four-chain structure comprising two light chains and two heavy chains. 
     The anti-PD-L1 antibody of the present invention may be prepared as described below. Briefly, an artificial gene is synthesized which comprises (i) the identified variable region sequences of a rat anti-bovine PD-L1 antibody and (ii) the constant region sequences of an antibody of an animal other than rat (e.g., canine or bovine) (preferably, human IgG4 antibody; antibody equivalent to human IgG4 antibody; or an immunoglobulin equivalent to human IgG1, in which mutations have been introduced into the relevant region to reduce ADCC activity and/or CDC activity). The resultant gene is inserted into a vector (e.g., plasmid), which is then introduced into a host cell (e.g., mammal cell such as CHO cell). The host cell is cultured, and the antibody of interest is collected from the resultant culture. 
     The amino acid sequence and the nucleotide sequence of the VL of the rat anti-bovine PD-L1 antibody identified by the present inventors are shown in SEQ ID NOS: 1 and 5, respectively. Further, the nucleotide sequence after codon optimization is shown in SEQ ID NO: 15. 
     The amino acid sequence and the nucleotide sequence of the VH of the rat anti-bovine PD-L1 antibody identified by the present inventors are shown in SEQ ID NOS: 2 and 6, respectively. Further, the nucleotide sequence after codon optimization is shown in SEQ ID NO: 16. 
     The amino acid sequence and the nucleotide sequence of the CL (Lambda chain, GenBank: E02824.1) of a canine antibody are shown in SEQ ID NOS: 3 and 7, respectively. Further, the nucleotide sequence after codon optimization is shown in SEQ ID NO: 17. 
     The amino acid sequence and the nucleotide sequence of the CL (Lambda chain, GenBank: X62917) of a bovine antibody are shown in SEQ ID NOS: 100 and 101, respectively. Further, the nucleotide sequence after codon optimization is shown in SEQ ID NO: 104. 
     The amino acid sequence and the nucleotide sequence of the CH (IgG-D chain, GenBank: AF354267.1) of the canine antibody are shown in SEQ ID NOS: 4 and 8, respectively. Further, the nucleotide sequence after codon optimization is shown in SEQ ID NO: 18. 
     The amino acid sequence and the nucleotide sequence (after codon optimization) of the CH (IgG1 chain, modified from GenBank: X62916) of the bovine antibody are shown in SEQ ID NOS: 102 and 103, respectively. 
     Further, SEQ ID NO: 9 shows the amino acid sequence of a chimeric light chain consisting of the VL of the rat anti-bovine PD-L1 antibody and the CL (Lambda chain, GenBank: E02824.1) of the canine antibody. The nucleotide sequence (after codon optimization) of the chimeric light chain consisting of the VL of the rat anti-bovine PD-L1 antibody and the CL (Lambda chain, GenBank: E02824.1) of the canine antibody is shown in SEQ ID NO: 19. 
     Further, SEQ ID NO: 105 shows the amino acid sequence of a chimeric light chain consisting of the VL of the rat anti-bovine PD-L1 antibody and the CL (Lambda chain, GenBank: X62917) of the bovine antibody. The nucleotide sequence (after codon optimization) of the chimeric light chain consisting of the VL of the rat anti-bovine PD-L1 antibody and the CL (Lambda chain, GenBank: X62917) of the bovine antibody is shown in SEQ ID NO: 107. 
     SEQ ID NO; 10 shows the amino acid sequence of a chimeric heavy chain consisting of the VH of the rat anti-bovine PD-L1 antibody and the CH (IgG-D chain, GenBank: AF354267.1) of the canine antibody. The nucleotide sequence (after codon optimization) of the chimeric heavy chain consisting of the VH of the rat anti-bovine PD-L1 antibody and the CH (IgG-D chain, GenBank: AF354267.1) of the canine antibody is shown in SEQ ID NO: 20. 
     SEQ ID NO: 106 shows the amino acid sequence of a chimeric heavy chain consisting of the VH of the rat anti-bovine PD-L1 antibody and the CH (IgG1 chain, modified from GenBank: X62916) of the bovine antibody. The nucleotide sequence (after codon optimization) of the chimeric heavy chain consisting of the VH of the rat anti-bovine PD-L1 antibody and the CH (IgG1 chain, modified from GenBank: X62916) of the bovine antibody is shown in SEQ ID NO: 108. 
     Amino acid sequences and nucleotide sequences of CLs and CHs for various animals other than the above may be obtained from known databases for use in the present invention. 
     Amino acid sequences and nucleotide sequences of CLs and CHs for canine, ovine, porcine, water buffalo, human and bovine are summarized in the table below. 
     Table. 
       
     
       
         
           
               
               
               
               
               
               
               
             
               
                 TABLE 
               
               
                   
               
               
                   
                   
                   
                   
                 GenBank 
                   
                   
               
               
                   
                   
                   
                   
                 Accession 
                 IMGT 
                   
               
               
                 Species 
                 Ig Domain 
                 Nucleotide Sequence 
                 Amino Acid Sequence 
                 No. 
                 Database 
                 Reference 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Canine 
                 Canine 
                 IgG-D 
                 GCCTCCACCACGGCCCCCTCG 
                 ASTTAPSVFPLAPSCGSTS 
                 AF354267 
                 http://ww  
                 Tang L, et al., 
               
               
                 (Scienti- 
                 Ig 
                   
                 GTTTTCCCACTGGCCCCCAGC 
                 GSTVALACLVSGYFPEPVT 
                   
                 w.imgt.or 
                 Vet. Immunol. 
               
               
                 fic 
                 heavy 
                   
                 TGCGGGTCCACTTCCGGCTCC 
                 VSWNSGSLTSGVHTFPSV 
                   
                 g/IMGTre 
                 Immunopathol, 
               
               
                 Name: 
                 chain 
                   
                 ACGGTGGCCCTGGCCTGCCTG 
                 LQSSGLYSLSSTVTVPSSR 
                   
                 pertoire/i 
                 80 (3-4), 259-270 
               
               
                 
                   Canis 
                 
                 constant 
                   
                 GTGTCAGGCTACTTCCCCGAG 
                 WPSETFTCNVVHPASNTK 
                   
                 ndex.php 
                 (2001), PMID: 
               
               
                 
                   lupus 
                 
                 region 
                   
                 CCTGTAACTGTGTCCTGGAAT 
                 VDKPVPKESTCKCISPCPV 
                   
                 ?section = 
                 11457479 
               
               
                   familiaris ) 
                 (CH1- 
                   
                 TCCGGCTCCTTGACCAGCGGT 
                 PESLGGPSVFIFPPKPKDIL 
                   
                 LocusGe 
                   
               
               
                   
                 CH3) 
                   
                 GTGCACACCTTCCCGTCCGTC 
                 RITRTPEITCVVLDLGRED 
                   
                 nes&amp;repe 
                   
               
               
                   
                   
                   
                 CTGCAGTCCTCAGGGCTCTAC 
                 PEVQISWFVDGKEVHTAK 
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                 TCCCTCAGCAGCACGGTGAC 
                 TQPREQQFNSTYRVVSVL 
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                 AGTGCCCTCCAGCAGGTGGC 
                 PIEHQDWLTGKEFKCRVN 
                   
                 pecies = do 
                   
               
               
                   
                   
                   
                 CCAGCGAGACCTTCACCTGCA 
                 HIGLPSPIERTISKARGQAH 
                   
                 g &amp; group = 
                   
               
               
                   
                   
                   
                 ACGTGGTCCACCCGGCCAGC 
                 QPSVYVLPPSPKELSSSDT 
                   
                 IGHC 
                   
               
               
                   
                   
                   
                 AACACTAAAGTAGACAAGCC 
                 VTLTCLIKDFFPPEIDVEW 
                   
                   
                   
               
               
                   
                   
                   
                 AGTGCCCAAAGAGTCCACCT 
                 QSNGQPEPESKYHTTAPQ 
                   
                   
                   
               
               
                   
                   
                   
                 GCAAGTGTATATCCCCATGCC 
                 LDEDGSYFLYSKLSVDKS 
                   
                   
                   
               
               
                   
                   
                   
                 CAGTCCCTGAATCACTGGGAG 
                 RWQQGDTFTCAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCTTCGGTCTTCATCTTTCC 
                 QNHYTDLSLSHSPGK* 
                   
                   
                   
               
               
                   
                   
                   
                 CCCGAAACCCAAGGACATCCT 
                 (SEQ ID NO: 4) 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGATTACCCGAACACCCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GATCACCTGTGTGGTGTTAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTGGGCCGTGAGGACCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTGCAGATCAGCTGGTTCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGATGGTAAGGAGGTGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCCAAGACGCAGCCTCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAGCAGCAGTTCAACAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTACCGTGTGGTCAGCGTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCCATTGAGCACCAGGACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCTCACCGGAAAGGAGTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTGCAGAGTCAACCACAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGCCTCCCGTCCCCCATCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGACTATCTCCAAAGCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGGCAAGCCCATCAGCCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGTGTATGTCCTGCCACCAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCAAAGGAGTTGTCATCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGACACGGTCACCCTGACCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTGATCAAAGACTTCTTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACCTGAGATTGATGTGGAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGAGCAATGGACAGCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCCCGAGAGCAAGTACCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGACTGCGCCCCAGCTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGACGGGTCCTACTTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTACAGCAAGCTCTCTGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAAGAGCCGCTGGCAGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAGACACCTTCACATGTGCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGATGCATGAAGCTCTACAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACCACTACACAGATCTATCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCTCCCATTCTCCGGGTAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGA (SEQ ID NO: 8) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Canine  
                 Ig 
                 CAGCCCAAGGCCTCCCCCT 
                 QPKASPSVTLFPPSSEE 
                 E02824 
                 Not 
                 None 
               
               
                   
                 Ig 
                 lambda 
                 CGGTCACACTCTTCCCGCC 
                 LGANKATLVCLISDFYP 
                   
                 registered 
                   
               
               
                   
                 light 
                 (CL) 
                 CTCCTCTGAGGAGCTCGGC 
                 SGVTVAWKASGSPVTQ 
                   
                   
                   
               
               
                   
                 chain 
                   
                 GCCAACAAGGCCACCCTGG 
                 GVETTKPSKQSNNKYA 
                   
                   
                   
               
               
                   
                 constant 
                   
                 TGTGCCTCATCAGCGACTTC 
                 ASSYLSLTPDKWKSHSS 
                   
                   
                   
               
               
                   
                 region 
                   
                 TACCCCAGCGGCGTGACGG 
                 FSCLVTHEGSTVEKKVA 
                   
                   
                   
               
               
                   
                   
                   
                 TGGCCTGGAAGGCAAGCGG 
                 PAECS* (SEQ ID NO: 3) 
                   
                   
                   
               
               
                   
                   
                   
                 CAGCCCCGTCACCCAGGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGAGACCACCAAGCCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAAGCAGAGCAACAACAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTACGCGGCCAGCAGCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGAGCCTGACGCCTGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTGGAAATCTCACAGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTTCAGCTGCCTGGTCACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGAGGGGAGCACCGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAAGAAGGTGGCCCCCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGTGCTCTTAG (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 7) 
                   
                   
                   
                   
               
               
                   
               
               
                 Ovine 
                 Ovine 
                 ‘IgG1 
                 GCCTCAACAACACCCCCGAA 
                 ASTTPPKVYPLTSCCGDTS 
                 X69797 
                 http://ww 
                 Dufour V. et al., 
               
               
                 (Scienti- 
                 heavy 
                   
                 AGTCTACCCTCTGACTTCTTG 
                 SSIVTLGCLVSSYMPEPVT 
                   
                 w.imgt.or 
                 J. Immunol., 156. 
               
               
                 tic 
                 chain 
                   
                 CTGCGGGGACACGTCCAGCT 
                 VTWNSGALTSGVHTFPAI 
                   
                 g/IMGTre 
                 2163-2170 
               
               
                 Name: 
                 constant 
                   
                 CCATCGTGACCCTGGGCTGCC 
                 LQSSGLYSLSSVVTVPAST 
                   
                 pertoir/li 
                 (1996). PMID: 
               
               
                 
                   Ovis 
                 
                 region 
                   
                 TGGTCTCCAGCTATATGCCCG 
                 SGAQTFICNVAHPASSTKV 
                   
                 ndex.php 
                 8690905 
               
               
                   aries ) 
                 (CH1 ~   
                   
                 AGCCGGTGACCGTGACCTGG 
                 DKRVEPGCPDPCKHCRCP 
                   
                 ?section =  
                   
               
               
                   
                 CH3) 
                   
                 AACTCTGGTGCCCTGACCAGC 
                 PPELPGGPSVFIFPPKPKDT 
                   
                 LocusGe 
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCGGCC 
                 LTISGTPEVTCVVVDVGQ 
                   
                 nes &amp; repe 
                   
               
               
                   
                   
                   
                 ATCCTGCAGTCCTCCGGGCTC 
                 DDPEVQFSWFVDNVEVRT 
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                 TACTCTCTCAGCAGCGTGGTG 
                 ARTKPREEQFNSTFRVVSA 
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCAGCACCTC 
                 LPIQHQDWTGGKEFKCKV 
                   
                 pecies = sh 
                   
               
               
                   
                   
                   
                 AGGAGCCCAGACCTTCATCTG 
                 HNEALPAPIVRTISRTKGQ 
                   
                 eep &amp; grou 
                   
               
               
                   
                   
                   
                 CAACGTAGCCCACCCGGCCA 
                 AREPQVYVLAPPQEELSK 
                   
                 p = IGHC 
                   
               
               
                   
                   
                   
                 GCAGCACCAAGGTGGACAAG 
                 STLSVTCLVTGFYPDYIAV 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGTTGAGCCCGGATGCCCG 
                 EWQKNGQPESEDKYGTT 
                   
                   
                   
               
               
                   
                   
                   
                 GACCCATGCAAACATTGCCGA 
                 TSQLDADGSYFLYSRLRV 
                   
                   
                   
               
               
                   
                   
                   
                 TGCCCACCCCCTGAGCTCCCC 
                 DKNSWQEGDTYACVVMH 
                   
                   
                   
               
               
                   
                   
                   
                 GGAGGACCGTCTGTCTTCATC 
                 EALHNHYTQKSISKPPGK* 
                   
                   
                   
               
               
                   
                   
                   
                 TTCCCACCGAAACCCAAGGA 
                 (SEQ ID NO: 42) 
                   
                   
                   
               
               
                   
                   
                   
                 CACCCTTACAATCTCTGGAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCGAGGTCACGTGTGTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGGACGTGGGCCAGGATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCGAGGTGCAGTTCTCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTTCGTGGACAACGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCGCACGGCCAGGACAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCGAGAGAGGAGCAGTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCACCTTCCGCGTGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGCCCTGCCCATCCAGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGACTGGACTGGAGGAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGTTCAAGTGCAAGGTCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACGAAGCCCTCCCGGCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCGTGAGGACCATCTCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACCAAAGGGCAGGCCCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCCGCAGGTGTACGTCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCCACCCCAGGAAGAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAAAGCACGCTCAGCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCACCTGCCTGGTCACCGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTACCCAGACTACATCGCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGAGTGGCAGAAAAATGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCCTGAGTCGGAGGACAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTACGGCACGACCACATCCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTGGACGCCGACGGCTCCTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTTCCTGTACAGCAGGCTCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGGACAAGAACAGCTGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGAAGGAGACACCTACGCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTGTGGTGATGCACGAGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGCACAACCACTACACACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGTCGATCTCTAAGCCTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGTAAATGA (SEQ ID NO: 43) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLTSCCGDTS 
                 X70983 
                   
                 Clarkson C.A. et 
               
               
                   
                   
                   
                 AGTCTACCCTCTGACTTCTTG 
                 SSSSIVTLGCLVSSYMPEP 
                   
                   
                 al., Mol. 
               
               
                   
                   
                   
                 CTGCGGGGACCACGTCCAGCT 
                 VTVTWNSGALTSGVHTFP 
                   
                   
                 Immunol., 30, 
               
               
                   
                   
                   
                 CCAGCTCCATCGTGACCCTGG 
                 AILQSSGLYSLSSVVTVPA 
                   
                   
                 1195-1204 
               
               
                   
                   
                   
                 GCTGCCTGGTCTCCAGCTATAT 
                 STSGAQTFICNVAHPASSA 
                   
                   
                 (1993). PMID: 
               
               
                   
                   
                   
                 GCCCGAGCCGGTGACCGTGA 
                 KVDKRVGISSDYSKCSKP 
                   
                   
                 8413324 
               
               
                   
                   
                   
                 CCTGGAACTCTGGTGCCCTGA 
                 PCVSRPSVFIFPPKPKDSL 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCGGCGTGCACACCTTCC 
                 MITGTPEVTCVVVDVGQG 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCCATCCTGCAGTCCTCCG 
                 DPEVQFSWFVDNVEVRTA 
                   
                   
                   
               
               
                   
                   
                   
                 GGCTCTACTCTCTCAGCAGCG 
                 RTKPREEQFNSTFRVVSAL 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTGACCGTGCCGGCCAGC 
                 PIQHDHWTGGKEFKCKV 
                   
                   
                   
               
               
                   
                   
                   
                 ACCTCAGGAGCCCAGACCTTC 
                 HSKGLPAPIVRTISRAKGQ 
                   
                   
                   
               
               
                   
                   
                   
                 ATCTGCAACGTAGCCCACCCG 
                 AREPQVYVLAPPQEELSK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGCAGCGCCAAGGTGGA 
                 STLSVTCLVTGFYPDYIAV 
                   
                   
                   
               
               
                   
                   
                   
                 CAAGCGTGTTGGGATCTCCAG 
                 EWQRARQPESEDKYGTTT 
                   
                   
                   
               
               
                   
                   
                   
                 TGACTACTCCAAGTGTTCTAA 
                 SQLDADGSYFLYSRLRVD 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGCCTTGCGTGAGCCGACC 
                 KSSWQRGDTYACVVMHE 
                   
                   
                   
               
               
                   
                   
                   
                 GTCTGTCTTCATCTTCCCCCCG 
                 ALHNHYTQKSISKPPGK* 
                   
                   
                   
               
               
                   
                   
                   
                 AAACCCAAGGACAGCCTCAT 
                 (SEQ ID NO: 44) 
                   
                   
                   
               
               
                   
                   
                   
                 GATCACAGGAACGCCCGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCACGTGTGTGGTGGTGGACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGGCCAGGGTGACCCCGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCAGTTCTCCTGGTTCGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACAACGTGGAGGTGCGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCCAGGACAAAGCCGAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGCAGTTCAACAGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTCCGCGTGGTCAGCGCCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCATCCAGCACGACCACTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTGGAGGAAAGGAGTTCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCAAGGTCCACAGCAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTCCCGGCCCCCATCGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACCATCTCCAGGGCCAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGCAGGCCCGGGAGCCGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGTACGTCCTGGCCCCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGAAGAGCTCAGCAAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGCTCAGCGICACCTGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCACCGGCTTCTACCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTACATCCICCGTGGAGTGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGAGCGCGGCAGCCTGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGGAGGACAAGTACGGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACCACATCCCAGCTGGACGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGACGGCTCCTACTTCCTGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAGGCTCAGGGTGGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCAGCTGGCAAAGAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACACCTACGCGTGTGTGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATGCACGAGGCTCTGCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACTACACACAGAACITCGATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTAAGCCTCCGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 45) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Ovine 
                 Ig 
                 CCATCCGTCTTCCTCTTCAAA 
                 PSVFLFKPSEEQLRTGTVS 
                 X54110 
                 Not 
                 Jenne C.N. et al., 
               
               
                   
                 light 
                 kappa 
                 CCATCTGAGGAACAGCTGAG 
                 VVCLVNDFYPKDINVKVK 
                   
                 registered 
                 Dev. Comp. 
               
               
                   
                 chain 
                 (CK) 
                 GACCGGAACTGTCTCTGTCGT 
                 VDGVTQNSNFQNSFTDQD 
                   
                   
                 Immunol. 30 (1- 
               
               
                   
                 constant 
                   
                 GTGCTTGGTGAATGATTTCTA 
                 SKKSTYSLSSTLTLSSSEY 
                   
                   
                 2), 165-174 
               
               
                   
                 region 
                   
                 CCCCAAAGATATCAATGTCAA 
                 QSHNAYACEVSHKSLPTA 
                   
                   
                 (2006). PMID: 
               
               
                   
                   
                   
                 GGTGAAAGTGGATGGGGTTA 
                 LVKSFNKNEC* (SEQ ID 
                   
                   
                 16083958 
               
               
                   
                   
                   
                 CCCAGAACAGCAACTTCCAG 
                 NO: 46) 
                   
                   
                   
               
               
                   
                   
                   
                 AACAGCTTCACAGACCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAGAAAAGCACCTACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTCAGCAGCACCCTGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGTCCAGCTCAGAGTACCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCCATAACGCCTATGCGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGTCAGCCACAAGAGCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCACCGCCCTCGTCAAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTTCAATAAGAATGAATGTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AG (SEQ ID NO: 47) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 Ig 
                 GGTCAGCCCAAGTCCGCACC 
                 GQPKSAPSVTLFPPSTEEL 
                 AY734681 
                   
                   
               
               
                   
                   
                 lambda 
                 CTCGGTCACCCTGTTCCCGCC 
                 STNKATVVCLINDFYPGS 
                   
                   
                   
               
               
                   
                   
                 (CL) 
                 TTCCACGGAGGAGCTCAGTAC 
                 VNVVWKADGSTINQNVK 
                   
                   
                   
               
               
                   
                   
                   
                 CAACAAGGCCACCGTGGTGT 
                 TTQASKQSNSKYAASSYL 
                   
                   
                   
               
               
                   
                   
                   
                 GTCTCATCAACGACTTCTACC 
                 TLTGSEWKSKSSYTCEVT 
                   
                   
                   
               
               
                   
                   
                   
                 CGGGTAGCGTGAACGTGGTCT 
                 HEGSTVTKTVKPSECS* 
                   
                   
                   
               
               
                   
                   
                   
                 GGAAGGCAGATGGCAGCACC 
                 (SEQ ID NO: 48) 
                   
                   
                   
               
               
                   
                   
                   
                 ATCAATCAGAACGTGAAGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCAGGCCTCCAAACAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAGCAAGTACGCGGCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGCTACCTGACCCTGACGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGCGAGTGGAAGTCTAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGTTACACCTGCGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGCACGAGGGGAGCACCGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGAAGACAGTGAAGCCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGAGTGTTCTTAG (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 49) 
                   
                   
                   
                   
               
               
                   
               
               
                 Porcine 
                 Porcine 
                 IgG1 a   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCGRDT 
                 U03781 
                 http://ww 
                 Butler J.E. et al., 
               
               
                 (Scienti- 
                 Ig 
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCASSYFPEPV 
                   
                 w.imgt.or 
                 Immunogenetics 
               
               
                 fic 
                 heavy 
                   
                 GGCAGGGACACGTCTGGCCC 
                 TMTWNSGALTSGVHTFPS 
                   
                 g/IMGTre 
                 61(3): 209-230 
               
               
                 Name: 
                 chain 
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 VLQPSGLYSLSSMVTVPAS 
                   
                 pertoire/i 
                 (2009). PMID: 
               
               
                 
                   Sus 
                 
                 constant 
                   
                 GGCCTCAAGCTACTFCCCCGA 
                 SLSSKSYTCNVNHPATTTK 
                   
                 ndex.php 
                 19048248 
               
               
                   scrofa ) 
                 region 
                   
                 GCCAGTGACCATGACCTGGA 
                 VDKRVGTKTKPPCPICPGC 
                   
                 ?section =  
                 Kacskovics I. et 
               
               
                   
                 (CH1- 
                   
                 ACTCGGGCGCCCTGACCAGT 
                 EVAGPSVFIFPPKPKDTLM 
                   
                 LocusGe 
                 al., J. Immunol. 
               
               
                   
                 CH3) 
                   
                 GGCGTGCATACCTTCCCATCC 
                 ISQTPEVTCVVVDVSKEH 
                   
                 nes &amp; repe 
                 153(8): 3565- 
               
               
                   
                   
                   
                 GTCCTGCAGCCGTCAGGGCTC 
                 AEVQFSWYVDGVEVHTA 
                   
                 rtoire = ge 
                 3573 (1994). 
               
               
                   
                   
                   
                 TACTCCCTCAGCAGCATGGTG 
                 ETRPKEEQFNSTYRVVSV 
                   
                 netable &amp; s 
                 PMID: 7930579 
               
               
                   
                   
                   
                 ACCGTGCCGGCCAGCAGCCT 
                 LPIQHQDWLKGKEFKCKV 
                   
                 pecies = Pi 
                   
               
               
                   
                   
                   
                 GTCCAGCAAGAGCTACACCT 
                 NNVDLPAPITRTISKAIGQS 
                   
                 g &amp; group = 
                   
               
               
                   
                   
                   
                 GCAATGTCAACCACCCGGCCA 
                 REPQVYTLPPPAEELSRSK 
                   
                 IGHC 
                   
               
               
                   
                   
                   
                 CCACCACCAAGGTGGACAAG 
                 VTVTCLVIGFYPPDIHVEW 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGTTGGAACAAAGACCAA 
                 KSNGQPEPEGNYRTTPPQ 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCATGTCCCATATGCCC 
                 QDVDGTFFLYSKLAVDKA 
                   
                   
                   
               
               
                   
                   
                   
                 AGGCTGTGAAGTGGCCGGGC 
                 RWDHGETFECAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 CCTCGGTCTTCATCTTCCCTCC 
                 HNHYTQKSISKTQGK* 
                   
                   
                   
               
               
                   
                   
                   
                 AAAACCCAAGGACACCCTCA 
                 (SEQ ID NO: 50) 
                   
                   
                   
               
               
                   
                   
                   
                 TGATCTCCCAGACCCCCGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCACGTGCGTGGTGGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCAGCAAGGAGCACGCCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTCCAGTTCTCCTGGTACGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACGGCGTAGAGGTGCACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGCCGAGACGAGACCAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGAGCAGTTCAACAGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTACCGTGTGGTCAGCGICCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCATCCAGCACCAGGACTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTGAAGGGGAAGGAGTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTGCAAGGTCAACAACGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACCTCCCAGCCCCCATCACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGACCATCTCCAAGGCTATA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGCAGAGCCGGGAGCCGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGTACACCCTGCCCCCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCGAGGAGCTGTCCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAAAGTCACCGTAACCTGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCATTGGCTTCTACCCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGACATCCATGTTGAGTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCAACGGACAGCCGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGAGGGCAAFTACCGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCGCCCCAGCAGGACGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACGGGACCTTCTTCCTGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAGCTCGCGGTGGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGCAAGATGGGACCATGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAACATTTGAGTGTGCGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATGCACGAGGCTCTGCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACTACACCCAGAAGTCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAAGACTCAGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 51) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG1 b   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCGRDV 
                 U03778 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCLASSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACGTGTCTGGCCCT 
                 TVTWNSGALTSGVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTGGCCTTGGGCTGCCTG 
                 VLQPSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 GCCTCAAGCTACTTCCCCGAG 
                 SLSSKSYTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGTGACCGTGACCTGGAA 
                 VDKRVGIHQPQTCPICPGC 
                   
                   
                   
               
               
                   
                   
                   
                 CTCGGGCGCCCTGACCAGTG 
                 EVAGPSVFIFPPKPKDTLM 
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGCACACCTTCCCATCCG 
                 ISQTEVTCVVVDVSKEH 
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGCAGCCGTCAGGGCTCT 
                 AEVQFSWYVDGVEVHTA 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCCCTCAGCAGCATGGTGA 
                 ETRPKEEQFNSTYRVVSC 
                   
                   
                   
               
               
                   
                   
                   
                 CCGTGCCGGCCAGCAGCCTGT 
                 LPIQHQDWLKGKEFKCKV 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCAAGAGCTACACCTGC 
                 NNVDLPAPITRTISKAIGQS 
                   
                   
                   
               
               
                   
                   
                   
                 AATGTCAACCACCCGGCCACC 
                 REPQVYTLPPPAEELSRSK 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCAAGGTGGACAAGCG 
                 VTLTCLVIGFYPPDIHVEW 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGAATACACCAGCCGCA 
                 KSNGQPEPENTYRTTPPQ 
                   
                   
                   
               
               
                   
                   
                   
                 AACATGTCCCATATGCCCAGG 
                 QDVDGTFFLYSKLAVDKA 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTGAAGTGGCCGGGCCCTC 
                 RWDHGDKFECAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCTTCATCTTCCCTCCAAA 
                 HNHYTQKSISKTQGK* 
                   
                   
                   
               
               
                   
                   
                   
                 ACCCAAGGACACCCTCATGAT 
                 (SEQ ID NO: 52) 
                   
                   
                   
               
               
                   
                   
                   
                 CTCCCAGACCCCCGAGGTCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCGTGGTGGTGGACGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAAGGAGCACGCCGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCTCCTGGTACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTAGAGGTGCACACGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGAGACGAGACCAAAGGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGTTCAACAGCACCTACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGTGGTCAGCGTCCTGCCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGCACCAGGACTGGCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGGGAAGGAGTTCAAGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGGTCAACAACGTAGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCAGCCCCCATCACGAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCTCCAAGGCTATAGGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCCGGGAGCCGCAGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACACCCTGCCCCCACCCGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGAGCTGTCCAGGAGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTCACGCTAACCTGCCTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATTGGCTTCTACCCACCTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATCCATGTTGAGTGGAAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACGGACAGCCGGAGCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAACACATACCGCACCACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCCAGCAGGACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGACCTTCTTCCTGTACAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAACTCGCGGTGGACAAGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGATGGGACCATGGAGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AATTTGAGTGTGCGGTGATGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGCTCTGCACAACCACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCAGAAGTCCATCTCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGACTCAGGGTAAATGA (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 53) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 a   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCSRDTS 
                 U03779 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 GPNVALGCLASSYFPEPVT 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGGGACACGTCTGGCCC 
                 VTWNSGALSSGVHTFPSV 
                   
                   
                   
               
               
                   
                   
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 LQPSGLYSLSSMVTVPASS 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCTCAAGCTACTTCCCCGA 
                 LSSKSYTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGTGACCGTGACCTGGA 
                 VDKRVGTKTKPPCPICPAC 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGCGCCCTGTCCAGTG 
                 ESPGPSVFIFPPKPKDTLMI 
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGCATACCTTCCCATCCG 
                 SRTPQVTCVVVDVSQENP 
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGCAGCCGTCAGGGCTCT 
                 EVQFSWYVDGVEVHTAQ 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCCCTCAGCAGCATGGTGA 
                 TRPKEEQFNSTYRVVSVLP 
                   
                   
                   
               
               
                   
                   
                   
                 CCGTGCCGGCCAGCAGCCTGT 
                 IQHQDWLNGKEFKCKVN 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCAAGAGCTACACCTGC 
                 NKDLPAPITRIISKAKGQT 
                   
                   
                   
               
               
                   
                   
                   
                 AATGTCAACCACCCGGCCACC 
                 REPQVYTLPPHAEELSRSK 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCAAGGTGGACAAGCG 
                 VSITCLVIGFYPPDIDVEW 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGAACAAAGACCAAAC 
                 QRNGQPEPEGNYRTTPPQ 
                   
                   
                   
               
               
                   
                   
                   
                 CACCATGTCCCATATGCCCAG 
                 QDVDGTYFLYSKFSVDKA 
                   
                   
                   
               
               
                   
                   
                   
                 CCTGTGAATCCCAGGGCCCT 
                 SWQGGGIFQCAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 CGGTCTTCATCTTCCCTCCAA 
                 HNHYTQKSISKTPGK* 
                   
                   
                   
               
               
                   
                   
                   
                 AACCCAAGGACACCCTCATGA 
                 (SEQ ID NO: 54) 
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCCGGACACCCCAGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGCGTGGTGGTTGATGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGGAGAACCCGGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCTCCTGGTACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTAGAGGTGCACACGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGACGAGGCCAAAGGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGTTCAACAGCACCTACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGGTCAGCGTCCTACCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGCACCAGGACTGGCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACGGGAAGGAGTTCAAGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGGTCAACAACAAAGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCAGCCCCCATCACAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCATCTCCAAGGCCAAAGGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGACCCGGGAGCCGCAGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACACCCTGCCCCCACACGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGAGCTGTCCAGGAGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTCAGCATAACCTGCCTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATTGGCTTCTACCCACCTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATCGATGTCGAGTGGCAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAACGGACAGCCGGAGCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGGCAATTACCGCACCACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCCAGCAGGACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGACCTACTTCCTGTACAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTTCTCGGTGGACAAGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCTGGCAGGGTGGAGGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TATTCCAGTGTGCGGTGATGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGCTCTGCACAACCACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCAGAAGTCTATCTCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGACTCCGGGTAAATGA (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 55) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 b   
                 GCCCCCAAGACGGCCCCATTG 
                 APKTAPLVYPLAPCGRDT 
                 U03780 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCLASSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACACGTCTGGCCC 
                 TVTWNSGALTSGVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 VLQPSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCTCAAGCTACTTCCCCGA 
                 SLSSKSYTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGTGACCGTGACCTGGA 
                 VDKRVGTKTKPPCPICPAC 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGCGCCCTGACCAGT 
                 ESPGPSVFIFPPKPKDTLMI 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCATACCTTCCCATCC 
                 SRTPQVTCVVVDVSQENP 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCAGCCGTCAGGGCTC 
                 EVQFSWYVDGVEVHTAQ 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCCCTCAGCAGCATGGTG 
                 TRPKEFQFNSTYRVVSVLP 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCAGCAGCCT 
                 IQHQDWLNGKEFKCKVN 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCAGCAAGAGCTACACCT 
                 NKDLPAPITRIISKAKGQT 
                   
                   
                   
               
               
                   
                   
                   
                 GCAATGTCAACCACCCGGCCA 
                 REPQVYTLPPHAEELSRSK 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCACCAAGGTGGACAAG 
                 VSITCLVIGFYPPDIDVEW 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGTTGGAACAAAGACCAA 
                 QRNGQPEPEGNYRTTPPQ 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCATGTCCCATATGCCC 
                 QDVDGTYFLYSKFSVDKA 
                   
                   
                   
               
               
                   
                   
                   
                 AGCCTGTGAATCGCCAGGGCC 
                 SWQGGGIFQCAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 CTCGGTCTTCATCTTCCCTCCA 
                 HNHYTQKSISKTPGK* 
                   
                   
                   
               
               
                   
                   
                   
                 AAACCCAAGGACACCCTCAT 
                 (SEQ ID NO: 56) 
                   
                   
                   
               
               
                   
                   
                   
                 GATCTCCCGGACACCCCAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGTGCGTGGTAGTTGATGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCCAGGAGAACCCGGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGTTCTCCTGGTACGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGGCGTAGAGGTGCACACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCAGACGAGGCCAAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAGCAGTTCAACAGCACCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCGCGTGGTCAGCGTCCTGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCCAGCACCAGGACTGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAACGGGAAGGAGTTCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCAAGGTCAACAACAAAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTCCCAGCCCCCATCACAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GATCATCTCCAAGGCCAAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGACCCGGGAGCCGCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTACACCCTGCCCCCACACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGAGGAGCTGTCCAGGAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGTCAGCATAACCTGCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCATTGGCTTCTACCCACCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACATCGATGTCGAGTGGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAAACGGACAGCCGGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGGGCAATTACCGCACCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCGCCCCAGCAGGACGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGGGACCTACTTCCTGTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAAGTTCTCGGTGGACAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCCAGCTGGCAGGGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCATATTCCAGTGTGCGGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCACGAGGCTCTGCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACTACACCCAGAAGTCTATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAAGACTCCGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 57) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG3 
                 GCCTACAACACAGCTCCATCG 
                 AYNTAPSVYPLAPCGRDV 
                 EU372658 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGT 
                 SDHNVALGCLVSSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACGTGTCTGATCAT 
                 TVTWNSGALSRVVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTGGCCTTGGGCTGCCTT 
                 VLQPSGLYSLSSMVIVAAS 
                   
                   
                   
               
               
                   
                   
                   
                 GTCTCAAGCTACTTCCCCGAG 
                 SLSTLSYTCNVYHPATNTK 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGTGACCGTGACCTGGAA 
                 VDKRVDIEPPTPICPEICSC 
                   
                   
                   
               
               
                   
                   
                   
                 CTCGGGTGCCCTGTCCAGAGT 
                 PAAEVLGAPSVFLFPPKPK 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGCATACCTTCCCATCCGT 
                 DILMISRTPKVTCVVVDVS 
                   
                   
                   
               
               
                   
                   
                   
                 CCTGCAGCCGTCAGGGCTCTA 
                 QEEAEVQFSWYVDGVQL 
                   
                   
                   
               
               
                   
                   
                   
                 CTCCCTCAGCAGCATGGTGAT 
                 YTAQTRPMEEQFNSTYRV 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGCGGCCAGCAGCCTGT 
                 VSVLPIQHQDWLKGKEFK 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCCTGAGCTACACGTGCA 
                 CKVNNKDLLSPITRTISKA 
                   
                   
                   
               
               
                   
                   
                   
                 ACGTCTACCACCCGGCCACCA 
                 TGPSRVPQVYTLPPAWEEL 
                   
                   
                   
               
               
                   
                   
                   
                 ACACCAAGGTGGACAAGCGT 
                 SKSKVSITCLCTGFYPPDI 
                   
                   
                   
               
               
                   
                   
                   
                 GTTGACATCGAACCCCCCACA 
                 DVEWQSNGQQEPEGNYR 
                   
                   
                   
               
               
                   
                   
                   
                 CCCATCTGTCCCGAAATTTGC 
                 TTPPQQDVDGTYFLYSKL 
                   
                   
                   
               
               
                   
                   
                   
                 TCATGCCCAGCTGCAGAGGTC 
                 AVDKVRWQRGDLFQCAV 
                   
                   
                   
               
               
                   
                   
                   
                 CTGGGAGCACCGTCGGTCTTC 
                 MHEALHNHYTQKSISKTQ 
                   
                   
                   
               
               
                   
                   
                   
                 CTCTTCCCTCCAAAACCCAAG 
                 GK (SEQ ID NO: 58) 
                   
                   
                   
               
               
                   
                   
                   
                 GACATCCTCATGATCTCCCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCAAGGTCACGTGCGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGGTGGACGTGAGCCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGGCTGAAGTCCAGTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGGTACGTGGACCCCGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTGTACACGGCCCAGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGCCAATGGAGGAGCAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAACAGCACCTACCGCGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAGCGTCCTGCCCATCCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCAGGACTGGCTGAAGGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGAGTTCAAGTGCAAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAACAAAGACCTCCTTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCATCACGAGGACCATCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGGCTACAGGGCCGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGTGCCGCAGGTGTACACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGCCCCCAGCCTGGGAAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTGTCCAAGAGCAAAGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCATAACCTGCCTGGTCACTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTTCTACCCACCTGACATCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATGTCGAGTGGCAGAGCAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACAACAAGAGCCAGAGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAATTACCGCACCACCCCGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCAGGACGTGGATGGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTACTTCCTGTACAGCAAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGCGGTGGACAAGGTCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCAGCGTGGAGACCTATTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTGTGCGGTGATGCACGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCTCTGCACAACCACTACAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGAAGTCCATCTCCAAGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAGGGTAAATGA (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 59) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG4 a   
                 ACCTTCCCATCCGTCCTGCAG 
                 TFPSVLQPSGLYSLSSMVT 
                 U03782 
                   
                   
               
               
                   
                   
                   
                 CCGTCAGGGCTCTACTCCCTC 
                 VPASSLSSKSYTCNVNHPA 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGCATGGTGACCGTGCC 
                 TTTKVDKRVGTKTKPPCPI 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCAGCAGCCTGTCCAGCA 
                 CPACEGPGPSAFIFPPKPK 
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCTACACCTGCAATGTCA 
                 DTLMISRTPKVTCVVVDV 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCCGGCCACCACCACC 
                 SQENPEVQFSWYVDGVE 
                   
                   
                   
               
               
                   
                   
                   
                 AAGGTGGACAAGCGTGTTGG 
                 VHTAQTRPKEEQFNSTYR 
                   
                   
                   
               
               
                   
                   
                   
                 AACAAAGACCAAACCACCAT 
                 VVSVLPIQHQDWLNGKEF 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCCATATGCCCAGCCTGTG 
                 KCKVNNKDLPAPITRIISK 
                   
                   
                   
               
               
                   
                   
                   
                 AAGGGCCCGGGCCCTCGGCC 
                 AKGQTREPQVYTLPPPTE 
                   
                   
                   
               
               
                   
                   
                   
                 TTCATCTTCCCTCCAAAACCC 
                 ELSRSKVTLTCLVTGFYPP 
                   
                   
                   
               
               
                   
                   
                   
                 AAGGACACCCTCATGATCTCC 
                 DIDVEWQRNGQPEPEGNY 
                   
                   
                   
               
               
                   
                   
                   
                 CGGACCCCCAAGGTCACGTG 
                 RTTPPQQDVDGTYFLYSK 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGTGGTAGATGTGAGCCA 
                 LAVDKASWQRGDTFQCA 
                   
                   
                   
               
               
                   
                   
                   
                 GGAGAACCCGGAGGTCCAGT 
                 VMHEALHNHYTQKSIFKT 
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCTGGTACGTGGACGGCG 
                 PGK* (SEQ ID NO: 60) 
                   
                   
                   
               
               
                   
                   
                   
                 TAGAGGTGCACACGGCCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGCCAAAGGAGGAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTTCAACAGCACCTACCGCGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTCAGCGTCCTGCCCATCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACCAGGACTGGCTGAACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAAGGAGTTCAAGTGCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCAACAACAAAGACCTCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCCCCCATCACAAGGATCAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCCAAGGCCAAAGGGCAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCGGGAGCCGCAGGTGTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCTGCCCCCACCCACCGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCTGTCCAGGAGCAAAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGCTAACCTGCCTGGTCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCTTCTACCCACCTGACAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGATGTCGAGTGGCAAAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGGACAGCCGGAGCCAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAATTACCGCACCACCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCAGCAGGACGTGGACGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACCTACTTCCTGTACAGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTCGCGGTGGACAAGGCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTGGCAGCGTGGAGACACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTCCAGTGTGCGGTGATGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGCTCTGCACAACCACTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCAGAAGTCCATCTTCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTCCGGGTAAATGA (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 61) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG4 b   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCGRDV 
                 EU372654 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCLASSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACGTGTCTGGCCCT 
                 TVTWNSGALTSGVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTGGCCTTGGGCTGCCTG 
                 VLQPSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 GCCTCAAGCTACTTCCCCGAG 
                 SLSSKSYTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGTGACCGTGACCTGGAA 
                 VDKRVGIHQPQTCPICPAC 
                   
                   
                   
               
               
                   
                   
                   
                 CTCGGGCGCCCTGACCAGTG 
                 EGPGPSAFIFPPKPKDTLM 
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGCACACCTTCCCATCCG 
                 ISRTPKVTCVVVDVSQEN 
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGCAGCCGTCAGGGCTCT 
                 PEVQFSWYVDGVEVHTA 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCCCTCAGCAGCATGGTGA 
                 QTRPKEEQFNSTYRVVSV 
                   
                   
                   
               
               
                   
                   
                   
                 CCGTGCCGGCCAGCAGCCTGT 
                 LLIQHQDWLNGKEFKCK 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCAAGAGCTACACCTGC 
                 VNNKDLPAPITRIISKAKG 
                   
                   
                   
               
               
                   
                   
                   
                 AATGTCAACCACCCGGCCACC 
                 QTREPQVYTLPPPTEELSR 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCAAGGTGGACAAGCG 
                 SKVTLTCLVTGFYPPDIDV 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGAATACACCAGCCGCA 
                 EWQRNGQPEPEGNYRTTP 
                   
                   
                   
               
               
                   
                   
                   
                 AACATGTCCCATAIGCCCAGC 
                 PQQDVDGTYFLYSKLAVD 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTGAAGGGCCCGGGCCCT 
                 KASWQRGDTFQCAVMHE 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCCTTCATCTTCCCTCCAA 
                 ALHNGYT (SEQ ID NO: 
                   
                   
                   
               
               
                   
                   
                   
                 AACCCAAGGACACCCTCATGA 
                 62) 
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCCGGACCCCCAAGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGCGTGGTGGTTGATGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGGAGAACCCGGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCTCCTGGTACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTAGAGGTGCACACGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGACGAGGCCAAAGGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGTTCAACAGCACCTACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGGTCAGCGTCCTGCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGCACCAGGACTGGCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACGGGAAGGAGTTCAAGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGGTCAACAACAAAGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCAGCCCCCATCACAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCATCTCCAAGGCCAAAGGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGACCCGGGAGCCGCAGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACACCCTGCCCCCACCCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGAGCTGTCCAGGAGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTCACGCTAACCTGCCTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACTGGCTTCTACCCACCTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATCGATGTCGAGTGGCAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAACGGACAGCCGGAGCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGGCAATTACCGCACCACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCCAGCAGGACGTGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGACCTACTTCCTGTACAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGCTCGCGGTGGACAAGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCTGGCAGCGTGGAGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATTCCAGTGTGCGGTGATGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGCTCTGCACAACCACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCC (SEQ ID NO: 63) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG5 a   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCSRDTS 
                 EU372657 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 GPNVALGCLVSSYFPEPVT 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGGGACACGTCTGGCCC 
                 VTWNSGALTSGVHTFPSV 
                   
                   
                   
               
               
                   
                   
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 LQPSGLYSLSSMVTVPAHS 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCTCAAGCTACTTCCCCGA 
                 LSSKRYTCNVNHPATKTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGTGACCGTGACCTGGA 
                 VDLCVGRPCPICPGCEVA 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGCGCCCTGACCAGT 
                 GPSVFIFPPKPKDILMISRT 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCATCC 
                 PEVTCVVVDVSKEHAEV 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCAGCCGTCAGGGCTC 
                 QFSWYVDGEEVHTAEFRP 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCCCTCAGCAGCATGGTG 
                 KEEQFNSTYRVVSVLPIQH 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCCACAGCTTG 
                 EDWLKGKEFECKVNNED 
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGCAAGCGCTATACGTGC 
                 LPGPITRTISKAKGVVRSP 
                   
                   
                   
               
               
                   
                   
                   
                 AATGTCAACCACCCAGCCACC 
                 EVYTLPPPAEELSKSIVTLT 
                   
                   
                   
               
               
                   
                   
                   
                 AAAACCAAGGTGGACCTGTG 
                 CLVKSIFP?FIHVEWKING 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGACGACCATGTCCCAT 
                 KPEPENAYRTTPPQEDEDR 
                   
                   
                   
               
               
                   
                   
                   
                 ATGCCCAGGCTGTGAAGTGGC 
                 TYFLYSKLAVDKARWDH 
                   
                   
                   
               
               
                   
                   
                   
                 CGGGCCCTCGGTCTTCATCTT 
                 GETFECAVMHEALHNHY 
                   
                   
                   
               
               
                   
                   
                   
                 CCCTCCAAAACCCAAGGACAT 
                 TQKSISKTQGK* (SEQ ID 
                   
                   
                   
               
               
                   
                   
                   
                 CCTCATGATCTCCCGGACCCC 
                 NO: 64) 
                   
                   
                   
               
               
                   
                   
                   
                 CGAGGTCACGTGCGTGGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGACGTCAGCAAGGAGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCGAGGTCCAGTTCTCCTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACGTGGACGGCGAAGAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACACGGCCGAGACGAGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAAGGAGGAGCAGTTCAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCACCTACCGCGTGGTCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCCCATCCAGCACGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACTGGCTGAAGGGGAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTTCGAGTGCAAGGTCAACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAAGACCTCCCAGGCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCACGAGGACCATCTCCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCAAAGGGGTGGTACGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCGGAGGTGTACACCCTGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCACCCGCCGAGGAGCTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAAGAGCATAGTCACGCTAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTGCCTGGTCAAAAGCATCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCGNCTTTCATCCATGTTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTGGAAAATCAACGGAAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGAGCCAGAGAACGCATAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCACCACCCCGCCTCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACGAGGACAGGACCTACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGTACAGCAAGCTCGCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGACAAGGCAAGATGGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATGGAGAAACATTTGAGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGGTGATGCACGAGGCTCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACAACCACTACACCCAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCATCTCCAAGACTCAGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TAAATGA (SEQ ID NO: 65) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG5 b   
                 GCCTACAACACAGCTCCATCG 
                 AYNTAPSVYPLAPCGRDV 
                 EU372656 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGT 
                 SDHNVALGCLVSSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACGTGTCTGATCAT 
                 TVTWNWGAQTSGVHTFP 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTGGCCTTGGGCTGCCTG 
                 SVLQPSGLYSLSSTVTVPA 
                   
                   
                   
               
               
                   
                   
                   
                 GTCTCAAGCTACTTCCCCGAG 
                 HSLSSKCFTCNVNHPATTT 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGTGACCGTGACCTGGAA 
                 KVDLCVGKKTKPRCPICP 
                   
                   
                   
               
               
                   
                   
                   
                 CTGGGGCGCCCAGACCAGTG 
                 GCEVAGPSVFIFPPKPKDIL 
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGCACACCTTCCCATCCG 
                 MISRTPEVTCVVVDVSKE 
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGCAGCCGTCAGGGCTCT 
                 HAEVQFSWYVDGEEVHT 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCCCTCAGCAGCACGGTG 
                 AETRPKEEQFNSTYRVVS 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCCACAGCTTG 
                 VLPIQHEDWLKGKEFECK 
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGCAAGTGCTTCACGTGC 
                 VNNEDLPGPITRTISKAKG 
                   
                   
                   
               
               
                   
                   
                   
                 AATGTCAACCACCCGGCCACC 
                 VVRSPEVYTLPPPAEELSK 
                   
                   
                   
               
               
                   
                   
                   
                 ACCACCAAGGTGGACCTGTG 
                 SIVTLTCLVKSFFPPFIHVE 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGAAAAAAGACCAAGC 
                 WKINGKPEPENAYRTTPP 
                   
                   
                   
               
               
                   
                   
                   
                 CTCGATGTCCCATATGCCCAG 
                 QEDEDGTYFLYSKFSVEK 
                   
                   
                   
               
               
                   
                   
                   
                 GCTGTGAAGTGGCCGGGCCC 
                 FRWHSGGIHCAVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 TCGGTCTTCATCTTCCCTCCA 
                 HNHYT (SEQ ID NO: 66) 
                   
                   
                   
               
               
                   
                   
                   
                 AAACCCAAGGACATCCTCATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCTCCCGGACCCCCGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGTGCGTGGTGGTGGACGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAGGAGCACGCCGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCAGTTCTCCTGGTACGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGGCGAAGAGGTGCACACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCGAGACGAGACCAAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAGCAGTTCAACAGCACTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCGCGTGGTCAGCGTCCTGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCCAGCACGAGGACTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGAAGGGGAAGGAGTTCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCAAGGTCAACAACGAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCTCCCAGGCCCCATCACGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACCATCTCCAAGGCCAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGGTGGTACGGAGCCCGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTGTACACCCTGCCCCCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCGAGGAGCTGTCCAAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCATAGTCACGCTAACCTGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCAAAAGCTTCTTCCCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTTTCATCCATGTTGAGTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAATCAACGGAAAACCAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGAGAACGCATACCGCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACCCCGCCCCAGGAGGACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGACGGGACCTACTTCCTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCAAGTTCTCGGTGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTTCAGGTGGCACAGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGCATCCACTGTGCGGTGAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGAGGCTCTGCACAACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTACACCC (SEQ ID NO: 67) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG6  
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCGRDT 
                 EU372655 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCLASSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACACGTCTGGCCC 
                 TLTWNSGALTSGVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 VLQPSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCTCAAGCTACTTCCCCGA 
                 SLSSKSYTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGTGACCCTGACCTGGA 
                 VDLCVGRPCPICPACEGPG 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGCGCCCTGACCAGT 
                 PSVFIFPPKPKDTLMISRTP 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCATACCTTCCCATCC 
                 QVTCVVVDVSQENPEVQF 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCAGCCGTCAGGGCTC 
                 SWYVDGVEVHTAQTRPK 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCCCTCAGCAGCATGGTG 
                 EAQFNSTYRVVSVLPIQHE 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCAGCAGCCT 
                 DWLKGKEFECKVNNKDL 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCAGCAAGAGCTACACCT 
                 PAPITRIISKAKGPSREPQV 
                   
                   
                   
               
               
                   
                   
                   
                 GCAATGTCAACCACCCGGCCA 
                 YTLSPSAEELSRSKVSITCL 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCACCAAGGTGGACCTG 
                 VTGFYPPDIDVEWKSNGQ 
                   
                   
                   
               
               
                   
                   
                   
                 TGTGTTGGACGACCATGTCCC 
                 PEPEGNYRTTPPQQDVDG 
                   
                   
                   
               
               
                   
                   
                   
                 ATATGCCCAGCCTGTGAAGGG 
                 TYFLYSKLAVDKASWQRG 
                   
                   
                   
               
               
                   
                   
                   
                 CCCGGGCCCTCGGTCTTCATC 
                 DPFQCAVMHEALHNHYT 
                   
                   
                   
               
               
                   
                   
                   
                 TTCCCTCCAAAACCCAAGGAC 
                 (SEQ ID NO: 68) 
                   
                   
                   
               
               
                   
                   
                   
                 ACCCTCATGATCTCCCGGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCAGGTCACGTGCGTGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTAGATGTGAGCCAGGAAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCGGAGGTCCAGTTCTCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTATGTGGACGGTGTAGAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACACGGCCCAGACGAGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAAGGAGGCGCAGTTCAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCACCTACCGTGTGGTCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCCCATCCAGCACGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACTGGCTGAAGGGGAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTTCGAGTGCAAGGTCAACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAAAGACCTCCAGCCCCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCACAAGGATCATCTCCAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAAAGGGCCGAGCCGGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGCAGGTGTACACCCTGTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCCGCCGAGGAGCTGTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGCAAAGTCAGCATAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGCCTGGTCACTGGCTTCTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCACCTGACATCGATGTCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGAAGAGCAACGGACAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGAGCCAGAGGGCAATTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCACCACCCCGCCCCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACGTGGACGGGACCTACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGTACAGCAAGCTCGCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGACAAGGCCAGCTGGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGAGACCCATTCCAGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGGTGATGCACGAGGCTCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACAACCACTACACCC (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 69) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG6 b   
                 GCCCCCAAGACGGCCCCATCG 
                 APKTAPSVYPLAPCGRDT 
                 EU372653 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGGCCCCCTGC 
                 SGPNVALGCLASSYFPEPV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGGACACGTCTGGCCC 
                 TVTWNSGALTSGVHTFPS 
                   
                   
                   
               
               
                   
                   
                   
                 TAACGTGGCCTTGGGCTGCCT 
                 VLQPSGLYSLSSTVTVPAR 
                   
                   
                   
               
               
                   
                   
                   
                 GGCCTCAAGCTACTTCCCCGA 
                 SSSRKCFTCNVNHPATTTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCAGTGACCGTGACCTGGA 
                 VDLCVGRPCPICPACEGN 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGCGCCCTGACCAGT 
                 GPSVFIFPPKPKDTLMISRT 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCATCC 
                 PEVTCVVVDVSQENPEVQ 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGCAGCCGTCAGGGCTC 
                 FSWYVDGEEVHTAETRPK 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCCCTCAGCAGCACGGTG 
                 EEQFNSTYRVVSVLPIQHQ 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCGGCCAGGAGCTC 
                 DWLKGKEFECKVNNKDL 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCAGAAAGTGCTTCACGTG 
                 PAPITRIISKAKGPSREPQV 
                   
                   
                   
               
               
                   
                   
                   
                 CAATGTCAACCACCCGGCCAC 
                 YTLSPSAEELSRSKVSITCL 
                   
                   
                   
               
               
                   
                   
                   
                 CACCACCAAGGTGGACCTGT 
                 VTGFYPPDIDVEWKSNGQ 
                   
                   
                   
               
               
                   
                   
                   
                 GTGTTGGACGACCATGTCCCA 
                 PEPEGNYRSTPPQEDEDG 
                   
                   
                   
               
               
                   
                   
                   
                 TATGCCCAGCCTGTGAAGGGA 
                 TYFLYSKLAVDKARLQSG 
                   
                   
                   
               
               
                   
                   
                   
                 ACGGGCCCTCGGTCTTCATCT 
                 GIHCAVMHEALHNHYTQ 
                   
                   
                   
               
               
                   
                   
                   
                 TCCCTCCAAAACCCAAGGAC 
                 KSISKT (SEQ ID NO: 70) 
                   
                   
                   
               
               
                   
                   
                   
                 ACCCTCATGATCTCCCGGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCGAGGTCACGTGCGTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTAGATGTGAGCCAGGAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCGGAGGTCCAGTTCTCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTACGTGGACGGCGAAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGCACACGGCCGAGACGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCAAAGGAGGAGCAGTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCACCTACCGTGTGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGTCCTGCCCATCCAGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGACTGGCTGAAGGGAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGTTCGAGTGCAAGGTCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAAAGACCTCCCAGCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCATCACAAGGATCATCTCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGCCAAAGGGCCGAGCCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCCGCAGGTGTACACCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCCCATCCGCCGAGGAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCAGGAGCAAAGTCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TAACCTGCCTGGTCACTGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTACCCACCTGACATCGATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGAGTGGAAGAGCAACGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCCGGAGCCAGAGGGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTACCGCTCCACCCCGCCCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAGGACGAGGACGGGGACCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTTCCTGTACAGCAAACTCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGTGGACAAGGCGAGGTTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGAGTGGAGGCATCCACTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGGTGATGCACGAGGCTCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACAACCACTACACCCAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCATCTCCAAGACT (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 71) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Porcine 
                 Ig 
                   
                   
                 FP312898 
                 http://ww 
                 Schwartz J.C. et 
               
               
                   
                 Ig 
                 kappa 
                   
                   
                   
                 w.imgt.or 
                 al., Immunogeneti 
               
               
                   
                 light 
                 (CK) 
                   
                   
                   
                 g/IMGTre 
                 cs, 64, 303-311 
               
               
                   
                 chain 
                 variant 
                   
                   
                   
                 pertoire/i 
                 (2012). PMID: 
               
               
                   
                 constant 
                 1 
                   
                   
                   
                 ndex.php 
                 22109540 
               
               
                   
                 region 
                   
                   
                   
                   
                 ?section =  
                   
               
               
                   
                   
                   
                   
                   
                   
                 LocusGe 
                   
               
               
                   
                   
                   
                   
                   
                   
                 nes &amp; repe 
                   
               
               
                   
                   
                   
                   
                   
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                   
                   
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                   
                   
                   
                 pecies = Pi 
                   
               
               
                   
                   
                   
                   
                   
                   
                 g &amp; group = 
                   
               
               
                   
                   
                   
                   
                   
                   
                 IGLC 
                   
               
               
                   
               
               
                   
                   
                 Ig 
                   
                   
                 CU694848 
                   
                   
               
               
                   
                   
                 kappa 
                   
                   
                   
                   
                   
               
               
                   
                   
                 (CK) 
                   
                   
                   
                   
                   
               
               
                   
                   
                 variant 
                   
                   
                   
                   
                   
               
               
                   
                   
                 2 
                   
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 Ig 
                   
                   
                 CU467669 
                 http://ww 
                   
               
               
                   
                   
                 lambda 
                   
                   
                   
                 w.imgt.or 
                   
               
               
                   
                   
                 (CL) 
                   
                   
                   
                 g/IMGTre 
                   
               
               
                   
                   
                 variant 
                   
                   
                   
                 pertoire/i 
                   
               
               
                   
                   
                 1 
                   
                   
                   
                 ndex.php 
                   
               
               
                   
                   
                   
                   
                   
                   
                 ?section =  
                   
               
               
                   
                   
                   
                   
                   
                   
                 LocusGe 
                   
               
               
                   
                   
                   
                   
                   
                   
                 nes &amp; repe 
                   
               
               
                   
                   
                   
                   
                   
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                   
                   
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                   
                   
                   
                 pecies = Pi 
                   
               
               
                   
                   
                   
                   
                   
                   
                 g &amp; group = 
                   
               
               
                   
                   
                   
                   
                   
                   
                 IGKC 
                   
               
               
                   
               
               
                   
                   
                 Ig 
                   
                   
                 CU467599 
                   
                   
               
               
                   
                   
                 lambda 
                   
                   
                   
                   
                   
               
               
                   
                   
                 (CK) 
                   
                   
                   
                   
                   
               
               
                   
                   
                 variant 
                   
                   
                   
                   
                   
               
               
                   
                   
                 2 
                   
                   
                   
                   
                   
               
               
                   
               
               
                 Water 
                 Water 
                 IgG1? 
                 GAGCGGCGTGCACACCTTCCC 
                 SGVHTFPAVLQSSGLYSLS 
                 NW_005690903 
                 Not 
                 None 
               
               
                 buffalo 
                 buffalo 
                   
                 GGCCGTCCTTCAGTCCTCCGG 
                 STVTAPASATKSQTFTCNV 
                   
                 registered 
                   
               
               
                 (Scienti- 
                 Ig 
                   
                 GCTCTACTCTCTCAGCAGCAC 
                 AHPASSTKVDKAVVPPCR 
                   
                   
                   
               
               
                 fic 
                 heavy 
                   
                 GGTGACCGCGCCCGCCAGCG 
                 PKPCDCCPPPELPGGPSVF 
                   
                   
                   
               
               
                 Name: 
                 chain 
                   
                 CCACAAAAAGCCAGACCTTC 
                 IFPPKPKDTLTISGTPEVTC 
                   
                   
                   
               
               
                 
                   Bubalus 
                 
                 constant 
                   
                 ACCTGCAACGTAGCCCACCCG 
                 VVVDVGHDDPEVKFSWF 
                   
                   
                   
               
               
                   bubalis ) 
                 region 
                   
                 GCCAGCAGCACCAAGGTGGA 
                 VDDVEVNTARTKPREEQF 
                   
                   
                   
               
               
                   
                 (CH1- 
                   
                 CAAGGCTGTTGTTCCCCCATG 
                 NSTYRVVSALPIQHNDWT 
                   
                   
                   
               
               
                   
                 CH3) 
                   
                 CAGACCCGAAACCCTGTGATTG 
                 GGKEFKCKVYNEGLPAPI 
                   
                   
                   
               
               
                   
                   
                   
                 CTGCCCACCCCCTGAGCTCCC 
                 VRTISRTKGQAREPQVYV 
                   
                   
                   
               
               
                   
                   
                   
                 CGGAGGACCCTCTGTCTTCAT 
                 LAPPQDELSKSTVSITCMV 
                   
                   
                   
               
               
                   
                   
                   
                 CTTCCCACCAAAACCCAAGG 
                 TGFYPDYIAVEWQKDGQP 
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCTCACAATCTCTGGAA 
                 ESEDKYGTTPPQLDSDGS 
                   
                   
                   
               
               
                   
                   
                   
                 CTCCTGAGGTCACGTGTGTGG 
                 YFLYSRLRVNKNSWQEGG 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTGGACGTGGGCCACGAT 
                 AYTCVVMHE (SEQ ID NO: 
                   
                   
                   
               
               
                   
                   
                   
                 GACCCCGAGGTGAAGTTCTCC 
                 72) 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTTCGTGGACGATGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTAAACACAGCCAGGACGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCAAGAGAGGAGCAGTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCACCTACCGCGTGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCGCCCTGCCCATCCAGCACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGACTGGACTGGAGGAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGTTCAAGTGCAAGGTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AATGAAGGCCTCCCAGCCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCGTGAGGACCATCTCCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCAAAGGGCAGGCCCGGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCGCAGGTGTACGTCCTGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCACCCCAGGACGAGCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAAAAGCACGGTCAGCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTTGCATGGTCACTGGCTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACCCAGACTACATCGCCGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGTGGCAGAAAGATGGGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTGAGTCAGAGGACAAATA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCACGACCCCGCCCCAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACAGCGATGGCTCCTACTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTGTACAGCAGGCTCAGGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAACAAGAACAGCTGGCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGAGGCGCCTACACGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTAGTGATGCATGAGGC (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 73) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2? 
                 GCCTCCATCACAGCCCCGAAA 
                 ASITAPKVYPLTSCRGETS 
                 NW_005766143 
                   
                   
               
               
                   
                   
                   
                 GTCTACCCTCTGACTTCTTGC 
                 SSTVTLGCLVSSYMPEPVT 
                   
                   
                   
               
               
                   
                   
                   
                 CGCGGGGAAACGTCCAGCTC 
                 VTWNSGALKSGVHTFPAV 
                   
                   
                   
               
               
                   
                   
                   
                 CACCGTGACCCTGGGCTGCCT 
                 LQSSGLYSLSSTVTAPASA 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCTCCAGCTACATGCCCGA 
                 TKSQTFTCNVAHPASSTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCGGTGACCGTGACCTGGA 
                 VDTAVGFSSDCCKFPKPC 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGTGCCCTGAAGAGC 
                 VRGPSVFIFPPKPKDTLMI 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCGGCC 
                 TGNPEVTCVVVDVGRDN 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTTCAGTCCTCTGGGCTC 
                 PEVQFSWFVGDVEVHTG 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCTCTCAGCAGCACGGTG 
                 RSKPREEQFNSTYRVVSTL 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGCGCCCGCCAGCGCCAC 
                 PIQHNDWTGGKEFKCKV 
                   
                   
                   
               
               
                   
                   
                   
                 AAAAAGCCAGACCTTCACCT 
                 NNKGLPAPIVRTISRTKGQ 
                   
                   
                   
               
               
                   
                   
                   
                 GCAACGTAGCCCACCCGGCC 
                 AREPQVYVLAPPQEELSK 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGCACCAAGGTGGACAC 
                 STVSVTCMVTGFYPDYIA 
                   
                   
                   
               
               
                   
                   
                   
                 GGCTGTTGGGTTCTCCAGTGA 
                 VEWHRDRQAESEDKYRT 
                   
                   
                   
               
               
                   
                   
                   
                 CTGCTGCAAGTTTCCTAAGCC 
                 TPPQLDSDGSYFLYSRLKV 
                   
                   
                   
               
               
                   
                   
                   
                 TTGTGTGAGGGGACCATCTGT 
                 NKNSWQEGGAYTCVVMH 
                   
                   
                   
               
               
                   
                   
                   
                 CTTCATCTTCCCGCCGAAACC 
                 E (SEQ ID NO: 74) 
                   
                   
                   
               
               
                   
                   
                   
                 CAAAGACACCCTGATGATCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAAATCCCGAGGTCACATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTGGTGGTGGACGTGGGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGATAACCCCGAGGTGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTCTCCTGGTTCGTGGGTGAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGAGGTGCACACGGGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCGAAGCCGAGAGAGGAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTTCAACAGCACCFACCGCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCAGCACCCTGCCCATCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCACAATGACTGGACTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAAAGGAGITCAAGTGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTCAACAACAAAGGCCTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCCCCCATCGTGAGGACCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCAGGACCAAAGGGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCGGGAGCCGCAGGTGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTCCTGGCCCCACCCCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCTCAGCAAAAGCACGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAGCGTCACTTGCATGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGCTTCTACCCAGACTACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGCCGTAGAGTGGCATAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCGGCAGGCTGAGTCGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACAAGTACCGCACGACCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCCAGCTGGACAGCGATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTCCTACTTCCTGTACAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCTCAAGGTGAACAAGAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCTGGCAAGAAGGAGGCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTACACGTGTGTAGTGATGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAGGC (SEQ ID NO: 75) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG3? 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLASSCGDTS 
                 NW_005784206 
                   
                   
               
               
                   
                   
                   
                 AGTCTACCCTCTGGCATCCAG 
                 SSTVTLGCLVSSYMPEPVT 
                   
                   
                   
               
               
                   
                   
                   
                 CTGCGGGGACACGTCCAGCT 
                 VTWNSGALKNGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCGTGACCCTGGGCTGCC 
                 VRQSSGLYSLSSMVTMPT 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCTCCAGCTACATGCCCG 
                 STAGTQTFTCNVAHPASST 
                   
                   
                   
               
               
                   
                   
                   
                 AGCCGGTGACCGTGACCTGG 
                 KVDTAVTARHPVPKTPET 
                   
                   
                   
               
               
                   
                   
                   
                 AACTCGGGTGCCCTGAAGAA 
                 PIHPVKPPTQEPRDEKTPC 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCGTGCACACCTTCCCGGC 
                 QCPKCPEPLGGLSVFIFPP 
                   
                   
                   
               
               
                   
                   
                   
                 CGTCCGGCAGTCCTCCGGGCT 
                 KPKDTLTISGTPEVTCVVV 
                   
                   
                   
               
               
                   
                   
                   
                 CTACTCTCTCAGCAGCATGGT 
                 DVGQDDPEVQFSWFVDD 
                   
                   
                   
               
               
                   
                   
                   
                 GACCATGCCCACCAGCACCGC 
                 VEVHTARMKPREEQFNST 
                   
                   
                   
               
               
                   
                   
                   
                 AGGAACCCAGACCTTCACCT 
                 YRVVSALPIQHQDWLREK 
                   
                   
                   
               
               
                   
                   
                   
                 GCAACGTAGCCCACCCGGCC 
                 EFKCKVNNKGLPAPIVRTI 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGCACCAAGGTGGACAC 
                 SRTKGQAREPQVYVLAPP 
                   
                   
                   
               
               
                   
                   
                   
                 GGCTGTCACTGCAAGGCATCC 
                 REELSKSTLSLTCLITGFYP 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCCCGAAGACACCAGAGA 
                 EEVDVEWQRNGQPESED 
                   
                   
                   
               
               
                   
                   
                   
                 CACCTATCCATCCTGTAAAAC 
                 KYHTTPPQLDADGSYFLY 
                   
                   
                   
               
               
                   
                   
                   
                 CCCCAACCCAGGAGCCCAGA 
                 SRLRVNRSSWQEGDHYTC 
                   
                   
                   
               
               
                   
                   
                   
                 GATGAAAAGACACCCTGCCA 
                 AVMHEALRNHYKEKPISR 
                   
                   
                   
               
               
                   
                   
                   
                 GTGTCCCAAATGCCCAGAACC 
                 SPGK* (SEQ ID NO: 76) 
                   
                   
                   
               
               
                   
                   
                   
                 TCTGGGAGGACTGTCTGTCTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATCTTCCCACCGAAACCCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACACCCTCACAATCTCTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACGCCCGAGGTCACGTGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTGGTGGACGTGGGCCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GATGACCCCGAAGTGCAGTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCTGGTTCGTGGATGACGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGGTGCACACAGCCAGGAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGCCAAGAGAGGAGCAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAACAGCACCTACCGCGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAGCGCCCTGCCCATCCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCAGGACTGGCTGCGGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGAGTTCAAGTGCAAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAACAAAGGCCTCCCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCCATCGTGAGGACCATCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGACCAAAGGGCAGGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGGAGCCACAGGTGTATGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGCCCCACCCCGGGAAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTCAGCAAAAGCACGCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTCACCTGCCTAATCACCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTTCTACCCAGAAGAGGTAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGTGGAGTGGCAGAGAAAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGCAGCCTGAGTCAGAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGTACCACACGACCCCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCAGCTGGACGCTGACGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCTACTTCCTGTACAGCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCAGGGTGAACAGGAGCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGCAGGAAGGAGACCACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACGTGTGCAGTGATGCATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGCTTTACGGAATCACTACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGAGAAGCCCATCTCGAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCGGGTAAATGA (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 77) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Water 
                 Ig 
                 CAGCCCAAGTCCGCACCCTCA 
                 QPKSAPSVTLFPPSTEELS 
                 NW_005690786 
                 Not 
                 None 
               
               
                   
                 buffalo 
                 lambda? 
                 GTCACCCTGTTCCCACCCTCC 
                 ANKATLVCLISDFYPGSMT 
                   
                 registered 
                   
               
               
                   
                 Ig 
                   
                 ACGGAGGAGCTCAGCGCCAA 
                 VARKADGSTITRNVETTR 
                   
                   
                   
               
               
                   
                 light 
                   
                 CAAGGCCACCCTGGTGTGTCT 
                 ASKQSNSKYAASSYLSLT 
                   
                   
                   
               
               
                   
                 chain 
                   
                 CATCAGCGACTTCTACCCGGG 
                 GSEWKSKGSYSCEVTHEG 
                   
                   
                   
               
               
                   
                 constant 
                   
                 TAGCATGACCGTGGCCAGGA 
                 STVTKTVKPSECS* (SEQ 
                   
                   
                   
               
               
                   
                 region 
                   
                 AGGCAGACGGCAGCACCATC 
                 ID NO: 78) 
                   
                   
                   
               
               
                   
                 (CL) 
                   
                 ACCCGGAACGTGGAGACCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGGGCCTCCAAACAGAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCAAGTACGCGGCCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCTACCTGAGCCTGACGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCGAGTGGAAATCGAAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGTTACAGCTGCGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGCACGAGGGGAGCACCGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACAAAGACAGTGAAGCCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGAGTGTTCTTAG (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 79) 
                   
                   
                   
                   
               
               
                   
               
               
                 Human 
                 Human 
                 IgG4 
                 GAGTCCAAATATGGTCCCCCA 
                 ESKYGPPCPSCPAPEFLGG 
                 K01316 
                 http://ww 
                 Ellison J. et al., 
               
               
                 (Scienti- 
                 Ig 
                 variant 
                 TGCCCATCATGCCCAGCACCT 
                 PSVFLFPPKPKDTLMISRT 
                   
                 w.imgt.or 
                 DNA, 1, 11-18 
               
               
                 fic 
                 heavy 
                 1 
                 GAGTTCCTGGGGGGACCATCA 
                 PEVTCVVVDVSQEDPEVQ 
                   
                 g/IMGTre 
                 (1981). PMID: 
               
               
                 Name: 
                 chain 
                   
                 GTCTTCCTGTTCCCCCCAAAA 
                 FNWYNDGVEVHNAKTKP 
                   
                 pertoire/i 
                 6299662 
               
               
                 
                   Homo 
                 
                 constant 
                   
                 CCCAAGGACACTCTCATGATC 
                 REEQFNSTYRVVSVLTVL 
                   
                 ndex.php 
                   
               
               
                   sapiens ) 
                 region 
                   
                 TCCCGGACCCCTGAGGTCACG 
                 HQDWLNGKEYKCKVSNK 
                   
                 ?section =  
                   
               
               
                   
                 (CH1- 
                   
                 TGCGTGGTGGTGGACGTGAG 
                 GLPSSIEKTISKAKGQPRE 
                   
                 LocusGe 
                   
               
               
                   
                 CH3) 
                   
                 CCAGGAAGACCCCGAGGTCC 
                 PQVYTLPPSQEEMTKNQV 
                   
                 nes &amp; repe 
                   
               
               
                   
                   
                   
                 AGTTCAACTGGTACGTGGATG 
                 SLTCLVKGFYPSDIAVEWE 
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                 GCGTGGAGGTGCATAATGCCA 
                 SNGQPENNYKTTPPVLDS 
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                 AGACAAAGCCGCGGGAGGAG 
                 DGSFFLYSRLTVDKSRWQ 
                   
                 pecies = hu 
                   
               
               
                   
                   
                   
                 CAGTTCAACAGCACGTACCGT 
                 EGNVFSCSVMHEALHNH 
                   
                 man &amp; gro 
                   
               
               
                   
                   
                   
                 GTGGTCAGCGTCCTCACCGTC 
                 YTQKSLSLSLGK* (SEQ ID 
                   
                 up = IGHC 
                   
               
               
                   
                   
                   
                 CTGCACCAGGACTGGCTGAA 
                 NO: 12) 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCAAGGAGTACAAGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCTCCAACAAAGGCCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGTCCTCCATCGAGAAAACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCTCCAAAGCCAAAGGGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCCGAGAGCCACAGGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCTGCCCCCATCCCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGATGACCAAGAACCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCAGCCTGACCTGCCTGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGCTTCTACCCCAGCGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCGCCGTGGAGTGGGAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAATGGGCAGCCGGAGAACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTACAAGACCACGCCTCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCTGGACTCCGACGGCTCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTTCCTCTACAGCAGGCTAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGTGGACAAGAGCAGGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGAGGGGAATGTCTTCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCTCCGTGATGCATGAGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGCACAACCACTACACACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGAGCCTCTCCCTGTCTCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGTAAATGA (SEQ ID NO: 14) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 1gG4 
                 GAGTCCAAATATGGTCCCCCG 
                 ESKYGPPCPSCPAPEFLGG 
                 AJ001563 
                   
                 Brusco A. et al., 
               
               
                   
                   
                 variant 
                 TGCCCATCATGCCCAGCACCT 
                 PSVFLFPPKPKDTLMISRT 
                   
                   
                 Eur. J. 
               
               
                   
                   
                 2 
                 GAGTTCCTGGGGGGACCATCA 
                 PEVTCVVVDVSQEDPEVQ 
                   
                   
                 Immunogenet., 
               
               
                   
                   
                   
                 GTCTTCCTGTTCCCCCCAAAA 
                 FNWYVDGVEVHNAKTKP 
                   
                   
                 25, 349-355 
               
               
                   
                   
                   
                 CCCAAGGACACTCTCATGATC 
                 REEQFNSTYRVVSVLTVV 
                   
                   
                 (1998). PMID: 
               
               
                   
                   
                   
                 TCCCGGACCCCTGAGGTCACG 
                 HQPWLNGKEYKCKVSNK 
                   
                   
                 9805657 
               
               
                   
                   
                   
                 TGCGTGGTGGTGGACGTGAG 
                 GLPSSIEKTISKAKGQPRE 
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGAAGACCCCGAGGTCC 
                 PQVYTLPPSQEEMTKNQV 
                   
                   
                   
               
               
                   
                   
                   
                 AGTTCAACTGGTACGTGGATG 
                 SLTCLVKGFYPSDIAVEWE 
                   
                   
                   
               
               
                   
                   
                   
                 GCGTGGAGGTGCATAATGCCA 
                 SNGQPENNYKTTPPVLDS 
                   
                   
                   
               
               
                   
                   
                   
                 AGACAAAGCCGCGGGAGGAG 
                 DGSFFLYSRLTVDKSRWQ 
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCAACAGCACGTACCGT 
                 EGNVFSCSVMHEALHNH 
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTCAGCGTCCTCACCGTC 
                 YTQKSLSLSEGK (SEQ ID 
                   
                   
                   
               
               
                   
                   
                   
                 GTGCACCAGGACTGGCTGAA 
                 NO: 80) 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCAAGGAGTACAAGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCTCCAACAAAGGCCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGTCCTCCATCGAGAAAACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCTCCAAAGCCAAAGGGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCCGAGAGCCACAGGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCTGCCCCCATCCCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGATGACCAAGAACCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCAGCCTGACCTGCCTGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGCTTCTACCCCAGCGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCGCCGTGGAGTGGGAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAATGGGCAGCCGGAGAACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTACAAGACCACGCCTCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCTGGACTCCGACGGCTCCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTTCCTCTACAGCAGGCTAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGTGGACAAGAGCAGGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGAGGGGAATGTCTTCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCTCCGTGATGCATGAGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGCACAACCACTACACCXA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGAGCCTCTCCCTGTCTCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGTAAATGA (SEQ ID NO: 81) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG4 
                 GCACCTGAGTTCCTGGGGGG 
                 APEFLGGSVFLFPPKPKD 
                 AJ001564 
                   
                   
               
               
                   
                   
                 variant 
                 ACCATCAGTCTTCCTGTTCCC 
                 TLMISKTPEVTCVVVDVS 
                   
                   
                   
               
               
                   
                   
                 3 
                 CCCAAAACCCAAGGACACTC 
                 QEDPEVQFNWYVDGVEV 
                   
                   
                   
               
               
                   
                   
                   
                 TCATGATCTCCCGGACCCCTG 
                 HNAKTKPREEQFNSTYRV 
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCACGTGCGTGGTGGTG 
                 VSVLTVLHQDWLNGKEY 
                   
                   
                   
               
               
                   
                   
                   
                 GACGTGAGCCAGGAAGACCC 
                 KCKVSNKGLPSSIEKTISK 
                   
                   
                   
               
               
                   
                   
                   
                 CGAGGTCCAGTTCAACTGGTA 
                 AKGQPREPQVYTLPPSQE 
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGATGGCGTGGAGGTGCA 
                 EMTKNQVSLTCLVKGFYP 
                   
                   
                   
               
               
                   
                   
                   
                 TAATGCCAAGACAAAGCCGC 
                 SDIAVEWESNGQPENNYK 
                   
                   
                   
               
               
                   
                   
                   
                 GGGAGGAGCAGTTCAACAGC 
                 TTPPVLDSDGSFFLYSKLT 
                   
                   
                   
               
               
                   
                   
                   
                 ACGTACCGTGTGGTCAGCGTC 
                 VDKSRWQEGNVFSCSVM 
                   
                   
                   
               
               
                   
                   
                   
                 CTCACCGTCCTGCACCAGGAC 
                 HEALHNHYTQKSLSLSLG 
                   
                   
                   
               
               
                   
                   
                   
                 TGGCTGAACGGCAAGGAGTA 
                 K* (SEQ ID NO: 82) 
                   
                   
                   
               
               
                   
                   
                   
                 CAAGTGCAAGGTCTCCAACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGCCTCCCGTCCTCCATCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAAAACCATCTCCAAAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGGCAGCCCCGAGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGGTGTACACCCTGCCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCCCAGGAGGAGATGACCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAACCAGGTCAGCCTGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCCTGGTCAAAGGCTTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCAGCGACATCGCCGTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGGAGAGCAATGGGCAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGAGAACAACTACAAGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGCCTCCCGTGCTGGACTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGGCTCCTTCTTCCTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCAAGCTCACCGTGGACAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCAGGTGGCAGGAGGGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGTCTTCTCATGCTCCGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCATGAGGCTCTGCACAACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACTACACGCAGAAGAGCCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCCTGTCTCTGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 83) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Human 
                 Ig 
                 ACTGTGGCTGCACCATCTGTC 
                 TVAAPSVFIFPPSDEQLKS 
                 X96754 
                 http://ww 
                 None 
               
               
                   
                 Ig 
                 kappa 
                 TTCATCTTCCCGCCATCTGATG 
                 GTASVVCLLNNFYPREAK 
                   
                 w.imgt.or 
                   
               
               
                   
                 light 
                 (CK) 
                 AGCAGTTGAAATCTGGAACTG 
                 VQWKVDNALQSGNSQES 
                   
                 g/IMGTre 
                   
               
               
                   
                 chain 
                   
                 CCTCTGTTGTGTGCCTGCTGA 
                 VTEQDSKDSTYSLSSTLTL 
                   
                 perioire/i 
                   
               
               
                   
                 constant 
                   
                 ATAACTTCTATCCCAGAGAGG 
                 SKADYEKHKVYACEVTH 
                   
                 ndex.php 
                   
               
               
                   
                 region 
                   
                 CCAAAGTACAGTGGAAGGTG 
                 QGLSSPVTKSFNRGEC* 
                   
                 ?section = 
                   
               
               
                   
                   
                   
                 GATAACGCCCTCCAATCGGGT 
                 (SEQ ID NO: 11) 
                   
                 LocusGe 
                   
               
               
                   
                   
                   
                 AACTCCCAGGAGAGTGYCAC 
                   
                   
                 nes &amp; repe 
                   
               
               
                   
                   
                   
                 AGAGCAGGACAGCAAGGACA 
                   
                   
                 rtoire = ge 
                   
               
               
                   
                   
                   
                 GCACCTACAGCCTCAGCAGC 
                   
                   
                 netable &amp; s 
                   
               
               
                   
                   
                   
                 ACCCTGACGCTGAGCAAAGC 
                   
                   
                 pecies = hu 
                   
               
               
                   
                   
                   
                 AGACTACGAGAAACACAAAG 
                   
                   
                 man &amp; gro 
                   
               
               
                   
                   
                   
                 TCTACGCCTGCGAAGTCACCC 
                   
                   
                 up = IGKC 
                   
               
               
                   
                   
                   
                 ATCAGGGCCTGAGCTCGCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCACAAAGAGCTTCAACAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAGAGTGTTAG (SEQ ID NO: 
                   
                   
                   
                   
               
               
                   
                   
                   
                 13) 
                   
                   
                   
                   
               
               
                   
               
               
                 Bovine 
                 Bovine 
                 IgG1 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLSSCCGDK 
                 X62916 
                 hap://ww 
                 Symons D.B. et 
               
               
                 (Scienti- 
                 Ig 
                 variant 
                 AGTCTACCCTCTGAGTTCTTG 
                 SSSTVTLGCLVSSYMPEPV 
                   
                 w.imgt.or 
                 al. J. 
               
               
                 fic 
                 heavy 
                 1 
                 CTGCGGGGACAAGTCCAGCT 
                 TVTWNSGALKSGVHTFPA 
                   
                 g/IMGTre 
                 Immunogenet., 
               
               
                 Name: 
                 chain 
                   
                 CCACCGTGACCCTGGGCTGCC 
                 VLQSSGLYSLSSMVTVPG 
                   
                 pertoire/i 
                 14, 273-283 
               
               
                 
                   Bos 
                 
                 constant 
                   
                 TGGTCTCCAGCTACATGCCCG 
                 STSGQTFTCNVAHPASSTK 
                   
                 ndex.php 
                 (1987). PMID: 
               
               
                   taurus ) 
                 region 
                   
                 AGCCGGTGACCGTGACCTGG 
                 VDKAVDPTCKPSPCDCCP 
                   
                 ?section = 
                 3141517 
               
               
                   
                 (CH1- 
                   
                 AACTCGGGTGCCCTGAAGAG 
                 PPELPGGPSVFIFPPKPKDT 
                   
                 LocusGe 
                 Symons D.B. et 
               
               
                   
                 CH3) 
                   
                 CGGCGTGCACACCTTCCCGGC 
                 LTISGTPEVTCVVVDVGH 
                   
                 nes &amp; repe 
                 al., Mol. 
               
               
                   
                   
                   
                 TGTCCTTCAGTCCTCCGGGCT 
                 DDPEVKFSWFVDDVEVNT 
                   
                 rtoire = ge 
                 Immunol., 26, 
               
               
                   
                   
                   
                 GTACTCTCTCAGCAGCATGGT 
                 ATTKPREEQFNSTYRVVS 
                   
                 netable &amp; s 
                 841-850 (1989). 
               
               
                   
                   
                   
                 GACCGTGCCCGGCAGCACCT 
                 ALRIQHQDWTGGKEFKC 
                   
                 pecies = bo 
                 PMID: 2513487 
               
               
                   
                   
                   
                 CAGGACAGACCTTCACCTGC 
                 KVHNEGLPARIVRTISRTK 
                   
                 vine &amp; gro 
                 Kacskovics I. and 
               
               
                   
                   
                   
                 AACGTAGCCCACCCGGCCAG 
                 GPAREPQVYVLAPPQEEL 
                   
                 up = IGHC 
                 Butler J.E., Mot 
               
               
                   
                   
                   
                 CAGCACCAAGGTGGACAAGG 
                 SKSTVSLTCMVTSFYPDYI 
                   
                   
                 immunol., 33, 
               
               
                   
                   
                   
                 CTGTTGATCCCACATGCAAAC 
                 AVEWQRNGQPESEDKYG 
                   
                   
                 189-195 (1996). 
               
               
                   
                   
                   
                 CATCACCCTGTGACTGTTGCC 
                 TTPPQLDADSSYFLYSKLR 
                   
                   
                 PMID: 8649440 
               
               
                   
                   
                   
                 CACCCCCTGAGCTCCCCGGAG 
                 VDRNSWQEGDTYTCVVM 
                   
                   
                 Rabbani H. et al., 
               
               
                   
                   
                   
                 GACCCTCTGTCTTCATCTTCCC 
                 HEALHNHYTQKSTSKSAG 
                   
                   
                 Immunogenetics, 
               
               
                   
                   
                   
                 ACCGAAACCCAAGGACACCC 
                 K (SEQ ID NO: 84) 
                   
                   
                 46, 326-331 
               
               
                   
                   
                   
                 TCACAATCTCGGGAACGCCCG 
                   
                   
                   
                 (1997). PMID: 
               
               
                   
                   
                   
                 AGGTCACGTGTGTGGTGGTG 
                   
                   
                   
                 9218535 
               
               
                   
                   
                   
                 GACGTGGGCCACGATGACCC 
                   
                   
                   
                 Saini S.S. et al., 
               
               
                   
                   
                   
                 CGAGGTGAAGTTCTCCTGGTT 
                   
                   
                   
                 Scand. J. 
               
               
                   
                   
                   
                 CGTGGACGACGTGGAGGTAA 
                   
                   
                   
                 Immunol. 65, 32- 
               
               
                   
                   
                   
                 ACACAGCCACGACGAAGCCG 
                   
                   
                   
                 8 (2007). PMID: 
               
               
                   
                   
                   
                 AGAGAGGAGCAGTTCAACAG 
                   
                   
                   
                 17212764 
               
               
                   
                   
                   
                 CACCTACCGCGTGGTCAGCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTGCGCATCCAGCACCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGACTGGAGGAAAGGAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAAGTGCAAGGTCCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGGCCTCCCGGCCCCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGAGGACCATCTCCAGGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGGCCGGCCCGGGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGGTGTATGTCCTGGCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCAGGAAGAGCTCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGCACGGTCAGCCTCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCATGGTCACCAGCTTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGACTACATCGCCGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCAGAGAAACGGGCAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAGTCGGAGGACAAGTACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGACCCCGCCCCAGCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGCCGACAGCTCCTACTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGTACAGCAAGCTCAGGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACAGGAACAGCTGGCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGAGACACCTACACGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTGATGCACGAGGCCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACAATCACTACACGCAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCACCTCTAAGTCTGCGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TAAATGA (SEQ ID NO: 92) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG1 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLSSCCGDK 
                 X16701 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGAGTTCTTG 
                 SSSTVTLGCLVSSYMPEPV 
                 (M25278) 
                   
                   
               
               
                   
                   
                 2 
                 CTGCGGGGACAAGTCCAGCT 
                 TVTWNSGALKSGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCGTGACCCTGGGCTGCC 
                 VLQSSGLYSLSSMVTVPG 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCTCCAGCTACATGCCCG 
                 STSGQTFTCNVAHPASSTK 
                   
                   
                   
               
               
                   
                   
                   
                 AGCCGGTGACCGTGACCTGG 
                 VDKAVDPTCKPSPCDCCP 
                   
                   
                   
               
               
                   
                   
                   
                 AACTCGGGTGCCCTGAAGAG 
                 PPELPGGPSVFIFPPKPKDT 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCGTGCACACCTTCCCGGC 
                 LTISGTPEVTCVVVDVGH 
                   
                   
                   
               
               
                   
                   
                   
                 CGTCCTTCAGTCCTCCGGGCT 
                 DDPEVKFSWFVDDVEVNT 
                   
                   
                   
               
               
                   
                   
                   
                 GTACTCTCTCAGCAGCATGGT 
                 ATTKPREEQFNSTYRVVS 
                   
                   
                   
               
               
                   
                   
                   
                 GACCGTGCCCGGCAGCACCT 
                 ALRIQHQDWTGGKEFKC 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGACAGACCTTCACCTGC 
                 KVHNEGLPAPIVRTISRTK 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTAGCCCACCCGGCCAG 
                 GPAREPQVYVLAPPQEEL 
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAAGGTGGACAAGG 
                 SKSTVSLTCMVTSFYPDYI 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTTGATCCCACATGCAAAC 
                 AVEWQRNGQPESEDKYG 
                   
                   
                   
               
               
                   
                   
                   
                 CATCACCCTGTGACTGTTGCC 
                 TTPPQLDADSSYFLYSKLR 
                   
                   
                   
               
               
                   
                   
                   
                 CACCCCCTGAGCTCCCCGGAG 
                 VDRNSWQEGDTYTCVVM 
                   
                   
                   
               
               
                   
                   
                   
                 GACCCTCTGTCTTCATCTTCCC 
                 HEALHNHYTQKSTSKSAG 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGAAACCCAAGGACACCC 
                 K* (SEQ ID NO: 85) 
                   
                   
                   
               
               
                   
                   
                   
                 TCACAATCTCGGGAACGCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCACGTGTGTGGTGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGTGGGCCACGATGACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGAGGTGAAGTTCTCCTGGTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGACGACGTGGAGGTAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACAGCCACGACGAAGCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGAGGAGCAGTTCAACAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACCTACCGCGTGGTCAGCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTGCGCATCCAGCACCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGACTGGAGGAAAGGAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAAGTGCAAGGTCCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGGCCTCCCGGCCCCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGAGGACCATCTCCAGGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGGCCGGCCCGGGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGGTGTATGTCCTGGCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCAGGAAGAGCTCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGCACGGTCAGCCATCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCATGGTCACCAGCTTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGACTACATCGCCGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCAGAGAAACGGGCAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAGTCGGAGGACAAGTACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGACCCCGCCCCAGCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGCCGACAGCTCCTACTTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGTACAGCAAGCTCAGGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACAGGAACAGCTGGCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGGAGACACCTACACGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTGATGCACGAGGCCCTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACAATCACTACACGCAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCACCTCTAAGTCTGCGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TAAATGA (SEQ ID NO: 93) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG1 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLSSCCGDK 
                 S82409 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGAGTTCTTG 
                 SSSTVTLGCLVSSYMPEPV 
                   
                   
                   
               
               
                   
                   
                 3 
                 CTGCGGGGACAAGTCCAGCT 
                 TVTWNSGALKSGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCGTGACCCTGGGCTGCC 
                 VLQSSGLYSLSSMVTVPG 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTCTCCAGCTACATGCCCG 
                 STSGTQTFTCNVAHPASST 
                   
                   
                   
               
               
                   
                   
                   
                 AGCCGGTGACCGTGACCTGG 
                 KVDKAVDPRCKTTCDCCP 
                   
                   
                   
               
               
                   
                   
                   
                 AACTCGGGTGCCCTGAAGAG 
                 PPELPGGPSVFIFPRKPKDT 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCGTGCACACCTTCCCGGC 
                 LTISGTPEVTCVVVDVGH 
                   
                   
                   
               
               
                   
                   
                   
                 CGTCCTTCAGTCCTCCGGGCT 
                 DDPEVKFSWFVDDVEVNT 
                   
                   
                   
               
               
                   
                   
                   
                 CTACTCTCTCAGCAGCATGGT 
                 ATTKPREEQFNSTYRVVS 
                   
                   
                   
               
               
                   
                   
                   
                 GACCGTGCCCGGCAGCACCT 
                 ALRIQHQDWTGGKEFKC 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGAACCCAGACCTTCACCT 
                 KVHNEGLPARIVRTISRTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCAACGTAGCCCACCCGGCC 
                 GPAREPQVYVLAPPQEEL 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGCACCAAGGTGGACAA 
                 SKSTVSLTCMVTSFYPDYI 
                   
                   
                   
               
               
                   
                   
                   
                 GGCTGTTGATCCCAGATGCAA 
                 AVEWQRNGQPESEDKYG 
                   
                   
                   
               
               
                   
                   
                   
                 AACAACCTGTGACTGTTGCCC 
                 TIPPQLDADGSYFLYSRLR 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGCCTGAGCTCCCTGGAG 
                 VDRNSWQEGDTYTCVVM 
                   
                   
                   
               
               
                   
                   
                   
                 GACCCTGTCTTCATCTTCCC 
                 HEALHNHYTQKSTSKSAG 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGAAACCCAAGGACACCC 
                 K* (SEQ ID NO: 86) 
                   
                   
                   
               
               
                   
                   
                   
                 TCACAATCTCGGGAACGCCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCACGTGTGTGGTGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACGTGGGCCACGATGACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGAGGTGAAGTTCTCCTGGTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGACGACGTGGAGGTAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACAGCCACGACGAAGCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGAGGAGCAGTTCAACAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACCTACCGCGTGGTCAGCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTGCGCATCCAGCACCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGACTGGAGGAAAGGAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAAGTGCAAGGTCCACAAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAAGGCCTCCCAGCCCCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGAGGACCATCTCCAGGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGGGCCGGCCCGGGAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGGTGTATGTCCTGGCCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCAGGAAGAGCTCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAAGCACGGTCAGCCTCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCATGGTCACCAGCTTCTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGACTACATCGCCGTGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCAGAGAAATGGGCAGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGAGTCAGAGGACAAGTACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGACCCCTCCCCAGCTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGCCGACGGCTCCTACTTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTACAGCAGGCTCAGGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACAGGAACAGCTGGCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGACACCTACACGTGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGTGATGCACGAGGCCCTGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAATCACTACACGCAGAAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCACCTCTAAGTCTGCGGGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AATGA (SEQ ID NO: 94) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLASSCGDTS 
                 S82407 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGGCATCCAG 
                 SSTVTLGCLVSSYMPEPVT 
                   
                   
                   
               
               
                   
                   
                 1 
                 CTGCGGAGACACATCCAGCTC 
                 VTWNSGALKSGVHTFPAV 
                   
                   
                   
               
               
                   
                   
                   
                 CACCGTGACCCTGGGCTGCCT 
                 LQSSGLYSLSSMVTVPASS 
                   
                   
                   
               
               
                   
                   
                   
                 GGTGTCCAGCTACATGCCCGA 
                 SGQTFTCNVAHPASSTKV 
                   
                   
                   
               
               
                   
                   
                   
                 GCCGGTGACCGTGACCTGGA 
                 DKAVGVSIDCSKCHNQPC 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGTGCCCTGAAGAGC 
                 VREPSVFIFPPKPKDTLMI 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCGGCT 
                 TGTPEVTCVVVNVGHDN 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTTCAGTCCTCCGGGCTC 
                 PEVQFSWFVDDVEVHTAR 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCTCTCAGCAGCATGGTG 
                 SKPREEQFNSTYRVVSALP 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCCGCCAGCAGCTC 
                 IQHQDWTGGKEFKCKVN 
                   
                   
                   
               
               
                   
                   
                   
                 AGGACAGACCTTCACCTGCA 
                 NKGLSAPIVRIISRSKGPAR 
                   
                   
                   
               
               
                   
                   
                   
                 ACGTAGCCCACCCGGCCAGC 
                 EPQVYVLDPPKEELSKSTL 
                   
                   
                   
               
               
                   
                   
                   
                 AGCACCAAGGTGGACAAGGC 
                 SVTCMVTGFYPEDVAVEW 
                   
                   
                   
               
               
                   
                   
                   
                 TGTTGGGGTCTCCATTGACTG 
                 QRNRQTESEDKYRTTPPQ 
                   
                   
                   
               
               
                   
                   
                   
                 CTCCAAGTGTCATAACCAGCC 
                 LDTDRSYFLYSKLRVDRN 
                   
                   
                   
               
               
                   
                   
                   
                 TTGCGTGAGGGAACCATCTGT 
                 SWQEGDAYTCVVMHEAL 
                   
                   
                   
               
               
                   
                   
                   
                 CTTCATCTTCCCACCGAAACC 
                 HNHYMQKSTSKSAGK* 
                   
                   
                   
               
               
                   
                   
                   
                 CAAAGACACCCTGATGATCAC 
                 (SEQ ID NO: 87) 
                   
                   
                   
               
               
                   
                   
                   
                 AGGAACGCCCGAGGTCACGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGTGGTGGTGAACGTGGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGATAACCCCGAGGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTTCTCCTGGTTCGTGGATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGAGGTGCACACGGCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGTCGAAGCCAAGAGAGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCAACAGCACGTACCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTCAGCGCCCTGCCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAGGACTGGACTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAAAGGAGTTCAAGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCAACAACAAAGGCCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGGCCCCCATCGTGAGGATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATCTCCAGGAGCAAAGGGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCCCGGGAGCCGCAGGTGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATGTCCTGGACCCACCCAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGAGCTCAGCAAAAGCACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCAGCGTCACCTGCATGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCGGCTTCTACCCAGAAGAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTAGCCGTGGAGTGGCAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAACCGGCAGACTGAGTCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGACAAGTACCGCACGACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGCCCCAGCTGGACACCGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGCTCCTACTTCCTGTACAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAAGCTCAGGGTGGACAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACAGCTGGCAGGAAGGAGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCTACACGTGTGTGGTGATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACGAGGCCCTGCACAATCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TACATGCAGAAGTCCACCTCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTCTGCGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 95) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLSSCCGDK 
                 M36946 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGAGTTCTTG 
                 SSSTVTLGCLVSSYMPEPV 
                 (X06703) 
                   
                   
               
               
                   
                   
                 2 
                 CTGCGGGGACAAGTCCAGCT 
                 TVTWNSGALKSGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CCACCGTGACCCTGGGCTGCC 
                 VLQSSGLYSLSSMVTVPG 
                   
                   
                   
               
               
                   
                   
                   
                 TGGTGTCCAGCTACATGCCCG 
                 STSGQTFTCNVAHPASSTK 
                   
                   
                   
               
               
                   
                   
                   
                 AGCCGGTGACCGTGACCTGG 
                 VDKAVGVSSDCSKPNNQ 
                   
                   
                   
               
               
                   
                   
                   
                 AACTCGGGTGCCCTGAAGAG 
                 HCVREPSVFIFPPKPKDTL 
                   
                   
                   
               
               
                   
                   
                   
                 CGGCGTGCACACCTTCCCGGC 
                 MITGTPEVTCVVVNVGHD 
                   
                   
                   
               
               
                   
                   
                   
                 CGTCCTTCAGTCCTCCGGGCT 
                 NPEVQFSWFVDDVEVHTA 
                   
                   
                   
               
               
                   
                   
                   
                 CTACTCTCTCAGCAGCATGGT 
                 RTKPREEQFNSTYRVVSA 
                   
                   
                   
               
               
                   
                   
                   
                 GACCGTGCCCGGCAGCACCT 
                 LPIQHQDWTGGKEFKCKV 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGACAGACCTTCACCTGC 
                 NIKGLSASINRIIRSKGPA 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTAGCCCACCCGGCCAG 
                 REPQVYVLDPPKEELSKS 
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAAGGTGGACAAGG 
                 TVSVTCMVIGFYPEDVDV 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTTGGGGTCTCCAGTGACT 
                 EWQRDRQTESEDKYRTTP 
                   
                   
                   
               
               
                   
                   
                   
                 GCTCCAAGCCTAATAACCAGC 
                 PQLDADRSYFLYSKLRVD 
                   
                   
                   
               
               
                   
                   
                   
                 ATTGCGTGAGGGAACCATCTG 
                 RNSWQRGDTYTCVVMHE 
                   
                   
                   
               
               
                   
                   
                   
                 TCTTCATCTTCCCACCGAAAC 
                 ALHNHYMQKSTSKSAGK 
                   
                   
                   
               
               
                   
                   
                   
                 CCAAAGACACCCTGATGATCA 
                 * (SEQ ID NO: 88) 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGAACGCCCGAGGTCACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTGTGGTGGTGAACGTGGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCACGATAACCCCGAGGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTTCTCCTGGTTCGTGGACGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGAGGTGCACACGGCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACGAAGCCGAGAGAGGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTCAACAGCACGTACCGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTCAGCGCCCTGCCCATC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAGGACTGGACTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAAAGGAGTTCAAGTGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTCAACATCAAAGGCCTCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGCCTCCATCGTGAGGATCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTCCAGGAGCAAAGGGCCG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCCGGGAGCCGCAGGTGTAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTGGACCCACCCAAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGCTCAGCAAAAGCACGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCAGCGTCACCTGCATGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCGGCTTCTACCCAGAAGATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TAGACGTGGAGTGGCAGAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACCGGCAGACTGAGTCGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGACAAGTACCGCACGACCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCCAGCTGGACGCCGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCTCCTACTTCCTGTACAGC  
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGCTCAGGGTGGACAGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCTGGCAGAGAGGAGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTACACGTGTGTGGTGATGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGAGGCCCTGCACAATCACT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACATGCAGAAGTCCACCTCTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGTCTGCGGGTAAATGA (SEQ 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ID NO: 96) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG2 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLSSCCGDK 
                 X16702 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGAGTTCTTG 
                 SSSGVTLGCLVSSYMPEPV 
                 (M25279) 
                   
                   
               
               
                   
                   
                 3 
                 CTGCGGGGACAAGTCCAGCT 
                 TVTWNSGALKSGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CGGGGGTGACCCTGGGCTGC 
                 VLQSSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 CTGGTCTCCAGCTACATGCCC 
                 SSGTQTFTCNVAHPASSTK 
                   
                   
                   
               
               
                   
                   
                   
                 GAGCCGGTGACCGTGACCTG 
                 VDKAVGVSSDCSKPNNQ 
                   
                   
                   
               
               
                   
                   
                   
                 GAACTCGGGTGCCCTGAAGA 
                 HCVREPSVFIFPPKPKDTL 
                   
                   
                   
               
               
                   
                   
                   
                 GCGGCGTGCACACCTTCCCGG 
                 MITGTPEVTCVVVNVGHD 
                   
                   
                   
               
               
                   
                   
                   
                 CCGTCCTTCAGTCCTCCGGGC 
                 NPEVQFSWFVDDVEVHTA 
                   
                   
                   
               
               
                   
                   
                   
                 TCTACTCTCTCAGCAGCATGG 
                 RTKPREEQFNSTYRVVSA 
                   
                   
                   
               
               
                   
                   
                   
                 TGACCGTGCCCGCCAGCAGCT 
                 LPIQHQDWTGGKEEKCKV 
                   
                   
                   
               
               
                   
                   
                   
                 CAGGAACCCAGACCTTCACCT 
                 NIKGLSASIVRIISRSKGPA 
                   
                   
                   
               
               
                   
                   
                   
                 GCAACGTAGCCCACCCGGCC 
                 REPQVYVLDPPKEELSKS 
                   
                   
                   
               
               
                   
                   
                   
                 AGCAGCACCAAGGTGGACAA 
                 TVSLTCMVIGFYPEDVDV 
                   
                   
                   
               
               
                   
                   
                   
                 GGCTGTTGGGGTCTCCAGTGA 
                 EWQRDRQTESEDKYRTTP 
                   
                   
                   
               
               
                   
                   
                   
                 CTGCTCCAAGCCTAATAACCA 
                 PQLDADRSYFLYSKLRVD 
                   
                   
                   
               
               
                   
                   
                   
                 GCATTGCGTGAGGGAACCATC 
                 RNSWQRGDTYTCVVMHE 
                   
                   
                   
               
               
                   
                   
                   
                 TGTCTTCATCTTCCCACCGAA 
                 ALHNHYMQKSTSKSAGK 
                   
                   
                   
               
               
                   
                   
                   
                 ACCCAAAGACACCCTGATGAT 
                 * (SEQ ID NO: 89) 
                   
                   
                   
               
               
                   
                   
                   
                 CACAGGAACGCCCGAGGTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGTGTGGTGGTGAACGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCCACGATAACCCCGAGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCAGTTCTCCTGGTTCGTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGACGTGGAGGTGCACACGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGACGAAGCCGAGAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGCAGTTCAACAGCACGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGCGTGGTCAGCGCCCTGCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CATCCAGCACCAGGACTGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGAGGAAAGGAGTTCAAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGCAAGGTCAACATCAAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTCTCGGCCTCCATCGTGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GATCATCTCCAGGAGCAAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCCGGCCCGGGAGCCGCAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGTATGTCCTGGACCCACCCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAAGAGCTCAGCAAAAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGGTCAGCCTCACCTGCATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTCATCGGCTTCTACCCAGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GATGTAGACGTGGAGTGGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGAGACCGGCAGACTGAGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGAGGACAAGTACCGCACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACCCCGCCCCAGCTGGACGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGACCGCTCCTACTTCCTGTA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAGCTCAGGGTGGACA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGAACAGCTGGCAGAGAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GACACCTACACGTGTGTGGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ATGCACGAGGCCCTGCACAAT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CACTACATGCAGAAGTCCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTAAGTCTGCGGGTAAATGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 (SEQ ID NO: 97) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG3 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLASSCGDTS 
                 U63638 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGGCATCCAG 
                 SSTVTLGCLVSSYMPEPVT 
                   
                   
                   
               
               
                   
                   
                 1 
                 CTGCGGAGACACATCCAGCTC 
                 VTWNSGALKSGVHTFPAV 
                   
                   
                   
               
               
                   
                   
                   
                 CACCGTGACCCTGGGCTGCCT 
                 RQSSGLYSLSSMVTVPASS 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCTCCAGCTACATGCCCGA 
                 SETQTFTCNVAHPASSTKV 
                   
                   
                   
               
               
                   
                   
                   
                 GCCGGTGACCCGTGACCTGGA 
                 DKAVTARRPVPTTPKTTIP 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGTGCCCTGAAGAGC 
                 PGKPTTPKSEVEKTPCQCS 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCGGCC 
                 KCPEPLGGLSVFIFPPKPK 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCGGCAGTCCTCTGGGCTG 
                 DILTISGTPEVTCVVVDV 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCTCTCAGCAGCATGGTG 
                 GQDDPEVQFSWFVDDVE 
                   
                   
                   
               
               
                   
                   
                   
                 ACTGTGCCCGCCAGCAGCTCA 
                 VHTARTKPREEQFNSTYR 
                   
                   
                   
               
               
                   
                   
                   
                 GAAACCCAGACCTTCACCTGC 
                 VVSALRIQHQDWLQGKEF 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTAGCCCACCCGGCCAG 
                 KCKVNNKGLPAPIVRTISR 
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAAGGTGGACAAGG 
                 TKGQAREPQVYVLAPPRE 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTCACTGCAAGGCGTCCAG 
                 ELSKSTLSLTCLITGFYPEE 
                   
                   
                   
               
               
                   
                   
                   
                 TCCCGACGACGCCAAAGACA 
                 IDVEWQRNGQPESEDKYH 
                   
                   
                   
               
               
                   
                   
                   
                 ACTATCCCTCCTGGAAAACCC 
                 TTAPQLDADGSYFLYSKL 
                   
                   
                   
               
               
                   
                   
                   
                 ACAACCCCAAAGTCTGAAGT 
                 RVNKSSWQEGDHYTCAV 
                   
                   
                   
               
               
                   
                   
                   
                 TGAAAAGACACCCTGCCAGT 
                 MHEALRNHYKEKSISRSP 
                   
                   
                   
               
               
                   
                   
                   
                 GTTCCAAATGCCCAGAACCTC 
                 GK* (SEQ ID NO: 90) 
                   
                   
                   
               
               
                   
                   
                   
                 TGGGAGGACTGTCTGTCTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTTCCCACCGAAACCCAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCTCACAATCTCGGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCGAGGTCACGTGTGTG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GTGGTGGACGTGGGCCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGACCCCGAGGTGCAGTTCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGTTCGTGGACGACGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTGCACACGGCCAGGACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGCCGAGAGAGGAGCAGTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAGCACCTACCGCGTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCGCCCTGCGCATCCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGACTGGCTGCAGGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGTTCAAGTGCAAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACAACAAAGGCCTCCCGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCATTGTGAGGACCATCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGACCAAAGGGCAGGCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGAGCCGCAGGTGTATGTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCCCCACCCCGGGAAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCAGCAAAAGCACGCTCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTCACCTGCCTGATCACCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTTCTACCCAGAAGAGATAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGAGTGGCAGAGAAATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGCCTGAGTCGGAGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTACCACACGACCGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCTGGAIGCTGACGGCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTACTTCCTGTACAGCAAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGGTGAACAAGAGCAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGAAGGAGACCACTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGTGTGCAGTGATGCACGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCTTTACGGAATCACTACAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGAGAAGTCCATCTCGAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCCGGGTAAATGA (SEQ ID 
                   
                   
                   
                   
               
               
                   
                   
                   
                 NO: 98) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                 IgG3 
                 GCCTCCACCACAGCCCCGAA 
                 ASTTAPKVYPLASRCGDT 
                 1163639 
                   
                   
               
               
                   
                   
                 variant 
                 AGTCTACCCTCTGGCATCCCG 
                 SSSTVTLGCLVSSYMPEPV 
                   
                   
                   
               
               
                   
                   
                 2 
                 CTGCGGAGACACATCCAGCTC 
                 TVTWNSGALKSGVHTFPA 
                   
                   
                   
               
               
                   
                   
                   
                 CACCGTGACCCTGGGCTGCCT 
                 VLQSSGLYSLSSMVTVPAS 
                   
                   
                   
               
               
                   
                   
                   
                 GGTCTCCAGCTACATGCCCGA 
                 TSETQTFTCNVAHPASSTK 
                   
                   
                   
               
               
                   
                   
                   
                 GCCGGTGACCGTGACCTGGA 
                 VDKAVTARRPVPTTPKTTI 
                   
                   
                   
               
               
                   
                   
                   
                 ACTCGGGTGCCCTGAAGAGT 
                 PPGKPTTQESEVEKTPCQC 
                   
                   
                   
               
               
                   
                   
                   
                 GGCGTGCACACCTTCCCGGCC 
                 SKCPEPLGGLSVFIFPPKP 
                   
                   
                   
               
               
                   
                   
                   
                 GTCCTTCAGTCCTCCGGGCTG 
                 KDTLTISGTPEVTCVVVD 
                   
                   
                   
               
               
                   
                   
                   
                 TACTCTCTCAGCAGCATGGTG 
                 VGQDDPEVQFSWFVDDV 
                   
                   
                   
               
               
                   
                   
                   
                 ACCGTGCCCGCCAGCACCTCA 
                 EVHTARTKPREEQFNSTY 
                   
                   
                   
               
               
                   
                   
                   
                 GAAACCCAGACCTTCACCTGC 
                 RVVSALRIQHQDWLQGKE 
                   
                   
                   
               
               
                   
                   
                   
                 AACGTAGCCCACCCGGCCAG 
                 FKCKVNNKGLPAPIVRTIS 
                   
                   
                   
               
               
                   
                   
                   
                 CAGCACCAAGGTGGACAAGG 
                 RTKGQAREPQVYVLAPPR 
                   
                   
                   
               
               
                   
                   
                   
                 CTGTCCTGCAAGGCGTCCAG 
                 EELSKSTLSLTCLITGFYPE 
                   
                   
                   
               
               
                   
                   
                   
                 TCCCGACGACGCCAAAGACA 
                 EIDVEWQRNGQPESEDKY 
                   
                   
                   
               
               
                   
                   
                   
                 ACCATCCCTCCTGGAAAACCC 
                 HTFAPQLDADGSYFLYSR 
                   
                   
                   
               
               
                   
                   
                   
                 ACAACCCAGGAGTCTGAAGT 
                 LRVNKSSWQEGDHYTCA 
                   
                   
                   
               
               
                   
                   
                   
                 TGAAAAGACACCCTGCCAGT 
                 VMHEALRNHYKEKSISRS 
                   
                   
                   
               
               
                   
                   
                   
                 GTTCCAAATGCCCAGAACCTC 
                 PGK* (SEQ ID NO: 91) 
                   
                   
                   
               
               
                   
                   
                   
                 TGGGAGGACTGTCTGTCTTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TCTTCCCACCGAAACCCAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACACCCTCACAATCTCGGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGCCCGAGGTCACGTGTGTG 
                   
                   
                   
                   
               
               
                   
               
               
                   
                   
                   
                 GTGGTGGACGTGGGCCAGGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGACCCCGAGGTGCAGTTCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTGGTTCGTGGACGACGTGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGTGCACACGGCCAGGACG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGCCGAGAGAGGAGCAGTT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAACAGCACCTACCGCGTGGT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCGCCCTGCGCATCCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGGACTGGCTGCAGGGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGGAGTTCAAGTGCAAGGTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AACAACAAAGGCCTCCCGGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCCCATTGTGAGGACCATCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGACCAAAGGGCAGGCCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGGAGCCCGCAGGTGTATGTCC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TGGCCCCACCCCGGGAAGAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTCAGCAAAAGCACGCTCAG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCTCACCTGCCTGATCACCGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 TTTCTACCCAGAAGAGATAGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGTGGAGTGGCAGAGAAATG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGCCTGAGTCGGAGGAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AAGTACCACACGACCGCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCAGCTGGATGCTGACGGCTC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CTACTTCCTGTACAGCAGGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGGGTGAACAAGAGCAGCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GGCAGGAAGGAGACCACTAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 ACGTGTGCAGTGATGCATGAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTTTACGGAATCACTACAAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGAAGTCCATCTCGAGGTCT 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CCGGGTAAATGA (SEQ ID NO: 
                   
                   
                   
                   
               
               
                   
                   
                   
                 99) 
                   
                   
                   
                   
               
               
                   
               
               
                   
                 Bovine, 
                 Ig 
                 CAGCCCAAGTCCCCACCCTCG 
                 QPKSPPSVTLFPPSTEELN 
                 X62917 
                 Not 
                 Chen L. et al., 
               
               
                   
                 Ig light 
                 lambda 
                 GTCACCCTGTTCCCGCCCTCC 
                 GNKATLVCLISDFYPGSVT 
                   
                 registered 
                 Vet. Immunol. 
               
               
                   
                 chain 
                   
                 ACGGAGGAGCTCAACGGCAA 
                 VVWKADGSTITRNVETTR 
                   
                   
                 Immunopathol, 
               
               
                   
                 constant 
                   
                 CAAGGCCACCCTGGTGTGTCT 
                 ASKQSNSKYAASSYLSLTS 
                   
                   
                 124, 284-294 
               
               
                   
                 region 
                   
                 CATCAGCGACTTCTACCCGGG 
                 SDWKSKGSYSCEVTHEGS 
                   
                   
                 (2008). PMID: 
               
               
                   
                 (CL) 
                   
                 TAGCGTGACCGTGGTCTGGAA 
                 TVTKTVKPSECS* (SEQ ID 
                   
                   
                 18538861 
               
               
                   
                   
                   
                 GGCAGACGGCAGCACCATCA 
                 NO: 100) 
                   
                   
                   
               
               
                   
                   
                   
                 CCCGCAACGTGGAGACCACC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGGGCCTCCAAACAGAGCAA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGCAAGTACGCGGCCAGCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCTACCTGAGCCTGACGAGC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 AGCGACTGGAAATCGAAAGG 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CAGTTACAGCTGCGAGGTCAC 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GCACGAGGGGAGCACCGTGA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 CGAAGACAGTGAAGCCCTCA 
                   
                   
                   
                   
               
               
                   
                   
                   
                 GAGTGTTCTTAG (SEQ ID NO: 
                   
                   
                   
                   
               
               
                   
                   
                   
                 101) 
               
               
                   
               
            
           
         
       
     
     The amino acid sequences as shown in SEQ ID NOS: 4, 3, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 12, 80, 82, 84-91, 100, 102 and 11 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) amino acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as the constant region of Ig heavy chain or light chain. 
     Although the constant region of wild-type human IgG1 has ADCC activity and CDC activity, it is known that these activities can be reduced by introducing amino acid substitutions and deletions into specific sites. In the case of animals other than human where the constant region of an immunoglobulin equivalent to human IgG4 has not been identified, mutations may be introduced into the relevant region of an immunoglobulin equivalent to human IgG1 so that the resultant constant region with reduced ADCC activity and CDC activity can be used. 
     The present invention provides an artificial genetic DNA comprising (a′) a DNA encoding a light chain comprising a light chain variable region (VL) containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the light chain constant region (CL) of an antibody of an animal other than rat and (b′) a DNA encoding a heavy chain comprising a heavy chain variable region (VH) containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the heavy chain constant region (CH) of an antibody of an animal other than rat. The present invention also provides a DNA encoding a light chain comprising a VL containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the CL of an antibody of an animal other than rat (i.e., the DNA of (a′) above). Further, the present invention also provides a DNA encoding a heavy chain comprising a VH containing CDR1 having GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the CH of an antibody of an animal other than rat (i.e., the DNA of (b′) above). 
     For (a) a light chain comprising a light chain variable region containing CDR1 having the amino acid sequence of QSLLYSENQKDY (SEQ ID NO: 37), CDR2 having the amino acid sequence of WAT and CDR3 having the amino acid sequence of GQYLVYPFT (SEQ ID NO: 38) and the light chain constant region of an antibody of an animal other than rat; and (b) a heavy chain comprising a heavy chain variable region containing CDR1 having the amino acid sequence of GYTFTSNF (SEQ ID NO: 39), CDR2 having the amino acid sequence of IYPEYGNT (SEQ ID NO: 40) and CDR3 having the amino acid sequence of ASEEAVISLVY (SEQ ID NO: 41) and the heavy chain constant region of an antibody of an animal other than rat, reference should be had to the foregoing description. The DNA of (a′) is a DNA (gene) encoding the light chain of (a); and the DNA of (b′) is a DNA (gene) encoding the heavy chain of (b). An artificial genetic DNA comprising the DNA of (a′) and the DNA of (′b) may be synthesized on commercial synthesizer. Restriction enzyme recognition sites, KOZAK sequences, poly-A addition signal sequences, promoter sequences, intron sequences or the like may be added to the artificial genetic DNA. 
     The present invention also provides a vector comprising the above-mentioned artificial genetic DNA. 
     As the vector,  Escherichia coli -derived plasmids (e.g., pBR322, pBR325, pUC12 or pUC13):  Bacillus subtilis -derived plasmids (e.g., pUB110, pTP5 or pC194), yeast-derived plasmids (e.g., pSH19 or pSH15); bacteriophages such as λ phage; animal viruses such as retrovirus or vaccinia virus, or insect pathogen viruses such as baculovirus may be used. In the Examples described later, pDC6 (Japanese Patent No. 5704753, U.S. Pat. No. 9,096,878, EU Patent 2385115, Hong Kong (China) patent HK1163739 and Australia Patent 2009331326) was used. 
     The vector may also comprise promoters, enhancers, splicing signals, poly-A addition signals, intron sequences, selection markers, SV40 replication origins, and so forth. 
     The present invention also provides a host cell transformed by the above vector. It is possible to prepare the anti-PD-L1 antibody of the invention by culturing the host cell and collecting the antibody of interest from the resultant culture. Therefore, the present invention also provides a method of preparing an antibody, comprising culturing the above-described host cell and collecting the anti-PD-L1 antibody of the invention from the culture. In the method of the present invention for preparing an antibody, a vector incorporating an artificial genetic DNA comprising a DNA encoding the light chain and a DNA encoding the heavy chain may be transfected into a host cell. Alternatively, a vector incorporating a DNA encoding the light chain and a vector incorporating a DNA encoding the heavy chain may be co-transfected into a host cell. 
     Examples of the host cell include, but are not limited to, bacterial cells (such as  Escherichia  bacteria,  Bacillus  bacteria or  Bacillus subtilis ), fungal cells (such as yeast or  Aspergillus ), insect cells (such as S2 cells or Sf cells), animal cells (such as CHO cells, COS cells, HeLa cells, C127 cells, 3T3 cells, BHK cells or HEK 293 cells) and plant cells. Among these, CHO-DG44 cell (CHO-DG44(dfhr − / − )) which is a dihydrofolate reductase deficient cell is preferable. 
     Introduction of a recombinant vector into a host cell may be performed by the methods disclosed in Molecular Cloning 2nd Edition, J. Sambrook et al., Cold Spring Harbor Lab. Press, 1989 (e.g., the calcium phosphate method, the DEAE-dextran method, transfection, microinjection, lipofection, electroporation, transduction, scrape loading, the shotgun method, etc.) or by infection. 
     The resultant transformant may be cultured in a medium, followed by collection of the anti-PD-L1 antibody of the present invention from the culture. When the antibody is secreted into the medium, the medium may be recovered, followed by isolation and purification of the antibody from the medium. When the antibody is produced within the transformed cells, the cells may be lysed, followed by isolation and purification of the antibody from the cell lysate. 
     Examples of the medium include, but are not limited to, OptiCHO medium, Dynamis medium, CD CHO medium, ActiCHO medium, FortiCHO medium, Ex-Cell CD CHO medium, BalanCD CHO medium. ProCHO 5 medium and Cellvento CHO-100 medium. 
     The pH of the medium varies depending on the cell to be cultured. Generally, a pH range from 6.8 to 7.6 is used; mostly, a pH range from 7.0 to 7.4 is appropriate. 
     When the cell to be cultured is CHO cells, culture may be performed by methods known to those skilled in the art. For example, it is usually possible to perform culturing in a gas-phase atmosphere having a CO 2  concentration of 0-40%, preferably 2-10%, at 30-39° C., preferably around 37° C. 
     The appropriate period of culture is usually from one day to three months, preferably from one day to three weeks. 
     Isolation and purification of the antibody may be performed by known methods. Known isolation/purification methods which may be used in the present invention include, but are not limited to, methods using difference in solubility (such as salting-out or solvent precipitation); methods using difference in molecular weight (such as dialysis, ultrafiltration, gel filtration or SDS-polyacrylamide gel electrophoresis); methods using difference in electric charge (such as ion exchange chromatography); methods using specific affinity (such as affinity chromatography); methods using difference in hydrophobicity (such as reversed phase high performance liquid chromatography); and methods using difference in isoelectric point (such as isoelectric focusing). 
     The anti-PD-L1 antibody of the present invention may be used as an antibody drug for animals or human. Therefore, the present invention provides a pharmaceutical composition comprising the above-described anti-PD-L1 antibody as an active ingredient. 
     The pharmaceutical composition of the present invention may be used for prevention and/or treatment of cancers and/or infections. Examples of cancers and/or infections include, but are not limited to, neoplastic diseases (e.g., malignant melanoma, lung cancer, gastric cancer, renal cancer, breast cancer, bladder cancer, esophageal cancer, ovarian cancer and the like), leukemia, Johne&#39;s disease, anaplasmosis, bacterial mastitis, mycotic mastitis, mycoplasma infections (such as mycoplasma mastitis, mycoplasma pneumonia or the like), tuberculosis,  Theileria orientalis  infection, cryptosporidiosis, coccidiosis, trypanosomiasis and leishmaniasis. 
     The anti-PD-L1 antibody of the present invention may be dissolved in buffers such as PBS, physiological saline or sterile water, optionally filter-sterilized with a filter or the like, and then administered to animal subjects (including human) by injection. To the solution of this antibody, additives (such as coloring agents, emulsifiers, suspending agents, surfactants, solubilizers, stabilizers, preservatives, antioxidants, buffers, isotonizing agents, pH adjusters and the like) may be added. As routes of administration, intravenous, intramuscular, intraperitoneal, subcutaneous or intradermal administration and the like may be selected. Transnasal or oral administration may also be used. 
     The dose and the number of times and frequency of administration of the anti-PD-L1 antibody of the present invention may vary depending on the symptoms, age and body weight of the animal subject, the method of administration, the dosage form and so on. For example, 0.1-100 mg/kg body weight, preferably 1-10 mg/kg body weight, per adult animal may usually be administered at least once, at such a frequency that enables confirmation of the desired effect. 
     While the pharmaceutical composition of the present invention may be used alone, it may be used in combination with surgical operations, radiation therapies, other immunotherapies such as cancer vaccine, or molecular target drugs. Synergistic effect can be expected from such combinations. 
     EXAMPLES 
     Hereinbelow, the present invention will be described in more detail with reference to the following Examples. However, the present invention is not limited to these Examples. 
     [Example 1] Rat-Canine Chimeric Anti-PD-L1 Antibody 
     1. Introduction 
     Programmed cell death 1 (PD-1), an immunoinhibitory receptor, and its ligand programmed cell death ligand 1 (PD-L1) are molecules identified by Prof. Tasuku Honjo et al., Kyoto University, as factors which inhibit excessive immune response and are deeply involved in immunotolerance. Recently, it has been elucidated that these molecules are also involved in immunosuppression in tumors. In the subject Example, for the purpose of establishing a novel therapy for canine neoplastic diseases, a chimeric antibody gene was prepared in which a variable region gene of a rat anti-bovine PD-L1 monoclonal antibody (4G12) capable of inhibiting the binding of canine PD-1 to PD-L1 was linked to a constant region gene of a canine immunoglobulin (IgG4). The resultant chimeric antibody gene was introduced into Chinese hamster ovary cells (CHO cells), which were cultured to produce a rat-canine chimeric anti-PD-L1 antibody c4G12. The effect of this chimeric antibody was confirmed in vitro and in vivo. 
     2. Materials and Methods 
     2.1 Bovine PD-L1 Monoclonal Antibody Producing Cells 
     The nucleotide sequence of bovine PD-L1 was identified (Ikebuchi R, Konnai S. Shirai T, Sunden Y, Murata S, Onuma M, Ohashi K. Vet Res. 2011 Sep. 26; 42:103). Based on the sequence information, a recombinant bovine PD-L1 was prepared. Rat was immunized with this recombinant protein to prepare a rat anti-bovine PD-L1 antibody (Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y, Murata S, Ohashi K. Immunology. 2014 August; 142(4):551-61; Clone 4G12 which would later serve as the variable region of the canine chimeric antibody of interest is described in this article.) 
     2.2 Identification of Full-Length Canine PD-1 and PD-L1 Genes 
     To determine the full lengths of canine PD-1 and PD-L1 cDNAs, PCR primers were first designed based on the putative nucleotide sequences of canine PD-1 and PD-L1 already registered at The National Center for Biotechnology Information (NCBI) (GenBank accession number: XM_543338 and XM_541302). Briefly, primers to amplify the inner sequence of the open reading frame (ORF) of each gene were designed (cPD-1 inner F and R, cPD-L1 inner F and R), and PCR was performed. For the amplified products, nucleotide sequences were determined with a capillary sequencer according to conventional methods. Further, to determine the nucleotide sequences of full-length PD-1 and PD-L1 cDNA, primers (cPD-1 5′ GSP and 3′ GSP; cPD-L1 5′ GSP and 3′GSP) were designed based on the canine PD-1 and PD-L1 cDNA sequences determined above. 5′-RACE and 3′-RACE were then performed using the 5′-RACE system for rapid amplification of cDNA ends and 3′-RACE system for rapid amplification of cDNA ends (Invitrogen), respectively. The resultant gene fragments of interest were sequenced as described (Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagawa Y, Nakajima C, Suzuki Y, Murata S, Ohashi K. PLoS One. 2014 Jun. 10; 9(6):e98415). 
     
       
         
           
               
               
            
               
                   
                 Primer (cPD-1 inner F): 
               
               
                   
                 (SEQ ID NO: 21) 
               
               
                   
                 AGGATGGCTCCTAGACTCCC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-1 inner R): 
               
               
                   
                 (SEQ ID NO: 22) 
               
               
                   
                 AGACGATGGTGGCATACTCG 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1 inner F): 
               
               
                   
                 (SEQ ID NO: 23) 
               
               
                   
                 ATGAGAATGTTTAGTGTCTT 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1 inner R): 
               
               
                   
                 (SEQ ID NO: 24) 
               
               
                   
                 TTATGTCTCTTCAAATTGTATATC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-1 5′GSP): 
               
               
                   
                 (SEQ ID NO: 25) 
               
               
                   
                 GTFGATCTGTGTGTTG 
               
               
                   
                   
               
               
                   
                 Primer (cPD-1 3′GSP): 
               
               
                   
                 (SEQ ID NO: 26) 
               
               
                   
                 CGGGACTTCCACATGAGCAT 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L3 5′GSP): 
               
               
                   
                 (SEQ ID NO: 27) 
               
               
                   
                 TTTTAGACAGAAAGTGA 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1 3′GSP): 
               
               
                   
                 (SEQ ID NO: 28) 
               
               
                   
                 GACCAGCTCTTCTTGGGGAA 
               
            
           
         
       
     
     2.3 Construction of Canine PD-1 and PD-L1 Expressing COS-7 Cells 
     For preparing canine PD-1-EGFP and PD-L1-EGFP expression plasmids, PCR was performed using a synthesized beagle PBMC-derived cDNA as a template and primers designed by adding XhoI and BamHI recognition sites (PD-1) and BglII and EcoRI recognition sites (PD-L1) on the 5′ side (cPD-1-EGFP F and R; cPD-L1-EGFP F and R). The resultant PCR products were digested with XhoI (Takara) and BamHI (Takara) (PD-1) and with BglII (New England Biolabs) and EcoRI (Takara) (PD-L1), and then purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics), followed by cloning into pEGFP-N2 vector (Clontech) treated with restriction enzymes in the same manner. The resultant expression plasmids of interest were extracted with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmids are designated as pEGFP-N2-cPD-1 and pEGFP-N2-cPD-L1. 
     
       
         
           
               
               
            
               
                   
                 Primer (cPD-1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 29) 
               
               
                   
                 CCGCTCGAGATGGGGAGCCGGCGGGGGCC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 30) 
               
               
                   
                 CGCGGATCCTGAGGGGCCACAGGCCGGGTC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 31) 
               
               
                   
                 GAAGATCTATGAGAATGTTTAGTGTC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 32) 
               
               
                   
                 GGAATTCTGTCTCTTCAAATTGTATATC 
               
            
           
         
       
     
     COS-7 cells were subcultured at a density of 5×10 4  cells/cm 2  in 6-well plates, and then cultured overnight in RPMI 1640 medium containing 10% inactivated fetal bovine serum and 0.01% L-glutamine at 37° C. in the presence of 5% CO 2 . The pEGFP-N2-cPD-1, pEGFP-N2-cPD-L1 or pEGFP-N2 (negative control) was introduced into COS-7 cells at 0.4 μg/cm 2  using Lipofectamine 2000 (Invitrogen). The cells were cultured for 48 hours (cPD-1-EGFP expressing cell and cPD-L1-EGFP expressing cell). In order to confirm the expression of canine PD-1 and PD-L1 in the thus prepared expressing cells, intracellular localization of enhanced green fluorescent protein (EGFP) was visualized with an inverted confocal laser microscope LSM700 (ZEISS) (Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagawa Y, Nakajima C, Suzuki Y, Murata S. Ohashi K. PLoS One. 2014 Jun. 10; 9(6):e98415). 
     2.4 Construction of Recombinant Canine PD-1, PD-L1 and CD80 
     In order to amplify the extracellular regions of canine PD-1, PD-L1 and CD80 estimated from their putative amino acid sequences, primers were designed. Briefly, primers having an NheI or EcoRV recognition sequence (PD-1 and PD-L1) added on the 5′ side (cPD-1-Ig F and R; cPD-L1-Ig F and R) or having an EcoRV or KpnI (CD80) recognition sequence added on the 5′ side (cCD80-Ig F and R) were designed. PCR was performed using a synthesized beagle PBMC-derived cDNA as a template. The PCR products were digested with NheI (Takara) and EcoRV (Takara) or with EcoRV (Takara) and KpnI (New England Biolabs) and purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics). The thus purified DNAs were individually cloned into pCXN2.1-Rabbit IgG Fc vector (Niwa et al., 1991; Zettimeissl et al., 1990; kindly provided by Dr. T. Yokomizo, Juntendo University Graduate School of Medicine, and modified in the inventors&#39; laboratory) treated with restriction enzymes in the same manner. The expression plasmids were purified with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmids are designated as pCXN2.1-cPD-1-Ig, pCXN2.1-cPD-L1-Ig and pCXN2.1-cCD80-Ig, respectively. 
     
       
         
           
               
               
            
               
                   
                 Primer (cPD-1-Ig F): 
               
               
                   
                 (SEQ ID NO: 33) 
               
               
                   
                 CCGCTCGAGATGGGGAGCCGGCGGGGGCC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-1-Ig R): 
               
               
                   
                 (SEQ ID NO: 34) 
               
               
                   
                 CGCGGATATCCAGCCCCTGCAACTGGCCGC 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1-Ig F): 
               
               
                   
                 (SEQ ID NO: 35) 
               
               
                   
                 CGCGGCTAGCATGAGAATGTTTAGTGTCTT 
               
               
                   
                   
               
               
                   
                 Primer (c-PD-L1-Ig R): 
               
               
                   
                 (SEQ ID NO: 36) 
               
               
                   
                 CGCGGATATCAGTCCTCTCACTTGCTGGAA 
               
               
                   
                   
               
               
                   
                 Primer (cCD80-Ig F): 
               
               
                   
                 (SEQ ID NO: 129) 
               
               
                   
                 CGCGGATATCATGGATTACACAGCGAAGTG 
               
               
                   
                   
               
               
                   
                 Primer (cCD80-Ig R): 
               
               
                   
                 (SEQ ID NO: 130) 
               
               
                   
                 CGGGGTACCCCAGAGCTGTTGCTGGTTAT 
               
            
           
         
       
     
     These expression vectors were individually transfected into Expi293F cells (Life Technologies) to obtain a culture supernatant containing a recombinant Ig fusion protein. The recombinant protein produced was purified from the supernatant with Ab Capcher Extra (Protein A mutant; ProteNova). After buffer exchange with phosphate-buffered physiological saline (PBS; pH 7.4) using PD-MidiTrap G-25 (GE Healthcare), each recombinant protein was stored at −30° C. until use in experiments (cPD-1-Ig, cPD-L1-Ig and cCD80-Ig). The concentration of each protein was measured with Pierce BCA Protein Assay Kit (Thermo Fisher Scientific) before use in subsequent experiments. 
     2.5 Identification of Rat Anti-Bovine PD-L1 Monoclonal Antibody Showing Cross-Reactivity with Canine PD-L1 
     In order to identify rat anti-bovine PD-L1 monoclonal antibody showing cross-reactivity with canine PD-L1, flow cytometry was performed using the anti-bovine PD-L1 antibody prepared in 2.1 above. The anti-bovine PD-L1 antibody (10 μg/ml) was reacted with 2×10 5 -1×10 6  cells at room temperature for 30 min. After washing, the anti-bovine PD-L1 antibody was detected with allophycocyanine-labeled anti-rat Ig goat antibody (Beckman Coulter). FACS Verse (Becton, Dickinson and Company) was used for analysis. As negative controls, rat IgG2a (κ) isotype control (BD Biosciences), rat IgG1 (κ) isotype control (BD Biosciences) and rat IgM (κ) isotype control (BD Biosciences) were used. For every washing operation and dilution of antibodies, 10% inactivated goat serum-supplemented PBS was used (Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagawa Y, Nakajima C, Suzuki Y, Murata S, Ohashi K. PLoS One. 2014 Jun. 10; 9(6):e98415 which is an article describing the use of three bovine PD-L1 monoclonal antibodies: 4G12 (Rat IgG2a (κ)), 5A2 (Rat IgG1 (κ)) and 6G7 (Rat IgM (κ)). 
     2.6 Selection Test of Variable Region for Establishment of Rat-Canine Chimeric Anti-PD-L1 Antibody 
     Out of 10 clones of rat anti-bovine PD-L1 monoclonal antibody which showed cross-reactivity with canine PD-L1, 4G12 (Rat IgG2a (κ)), 5A2 (Rat IgG1 (κ)) and 6G7 (Rat IgM (κ)) were selected and check was made to see whether these antibodies would inhibit canine PD-1/PD-L1 binding. Briefly, canine PD-1-Ig (prepared in 2.4 above) was immobilized on flat bottomed 96-well plates and blocked with 1% BSA and 0.05% Tween 20-containing PBS. Canine PD-L1-Ig (prepared in 2.4 above) was biotinylated using Lightning-Link Biotin Conjugation Kit (Innova Bioscience) and reacted with various concentrations (0, 2.5, 5 and 10 μg/ml) of rat anti-bovine PD-L1 antibodies 4G12, 5A2 and 6G7 at 37° C. for 30 min, followed by addition to the 96-well plates. The binding of cPD-L1-Ig to cPD-1-Ig was measured by color reaction using Neutravidin-HRP (Thermo Fisher Scientific) and TMB one component substrate (Bethyl Laboratories). As a result, rat anti-bovine PD-L1 monoclonal antibodies 4G12 and 607 showed a good inhibitory activity against canine PD-1/PD-L1 binding, whereas 5A2 showed no binding inhibitory activity ( FIG. 1 ). 
     2.7 Preparation of Rat-Canine Chimeric Anti-PD-L1 Antibody Expressing Vector (FIG.  2 ) 
     Using rat anti-bovine PD-L1 monoclonal antibodies 4G12 and 6G7 which showed a good inhibitory activity against canine PD-1/PD-L1 binding ( FIG. 1 ) as the variable region, two types of rat-canine chimeric anti-PD-L1 antibodies were established. 
     Briefly, heavy chain and light chain variable region genes were identified from hybridomas producing rat anti-bovine PD-L1 monoclonal antibodies 4G12 and 6G7. Further, the heavy chain and light chain variable region genes of the above rat antibodies were linked to the constant region of heavy chain IgG4 and the constant region of light chain Lambda of a known canine antibody, respectively, to prepare nucleotide sequences, followed by codon optimization (SEQ ID NOS: 9 and 10 (amino acid sequences), SEQ ID NOS: 19 and 20 (nucleotide sequences after codon optimization). Then, synthesis of genes was performed so that NotI restriction enzyme recognition site, KOZAK sequence, chimeric antibody&#39;s light chain sequence, poly-A addition signal sequence (PABGH), promoter sequence (PCMV). SacI restriction enzyme recognition site, intron sequence (INRBG), KOZAK sequence, chimeric antibody&#39;s heavy chain sequence and XbaI restriction enzyme recognition site would be located in this order. The synthesized gene strands were individually incorporated into the cloning site (NotI and XbaI restriction enzyme recognition sequences downstream of PCMV and between INRBG and PABGH) of expression vector pDC6 (kindly provided by Prof. S. Suzuki. Hokkaido University Research Center for Zoonosis Control) using restriction enzyme recognition sequences so that the above-listed sequences would be located in the above-mentioned order ( FIG. 2 ). Thus, rat-canine chimeric anti-PD-L1 antibody expressing vectors were constructed. Each of the expression vectors was transfected into Expi293F cells (Life Technologies) to obtain a culture supernatant containing a chimeric antibody. The chimeric antibody was purified from the supernatant with Ab Capcher Extra (Protein A mutant; ProteNova) and further purified by gel filtration chromatography. SDS-PAGE was performed under non-reducing conditions using 10% acrylamide gel. Bands were stained with Quick-CBB kit (Wako Pure Chemical) and decolorized in distilled water. Although contaminant proteins were observed after protein A purification alone, a highly purified antibody could be obtained by performing gel filtration chromatography ( FIG. 3 ). It was confirmed by flow cytometry that the resultant purified antibodies specifically bound to canine PD-L1 expressing cells (data not shown). When the inhibitory activity of the two chimeric antibodies against canine PD-1/PD-L1 binding was examined by the method described in 2.6 above, rat-canine chimeric anti-PD-L1 antibody c4G12 showed a binding inhibitory activity similar to that of its original rat anti-bovine PD-L1 monoclonal antibody 4G12, whereas binding inhibition capacity was clearly attenuated in rat-canine chimeric anti-PD-L1 antibody c6G7 ( FIG. 4 ) Therefore, rat-canine chimeric anti-PD-L1 antibody c4G12 was selected as a therapeutic antibody, which incorporated the variable region sequences of rat anti-bovine PD-L1 monoclonal antibody 4G12 (SEQ ID NOS: 2 and 1 (amino acid sequences) and SEQ ID NOS: 16 and 15 (nucleotide sequences after codon optimization)). The amino acid sequence and the nucleotide sequence (after codon optimization) of the light chain of c4G12 are shown in SEQ ID NOS: 9 and 19, and the amino acid sequence and the nucleotide sequence (after codon optimization) of the heavy chain of c4G12 are shown in SEQ ID NOS: 10 and 20. 
     2.8 Expression of Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 
     Rat-canine chimeric anti-PD-L1 antibody c4G12 expressing vector pDC6 as used in 2.7 above was transfected into CHO-DG44 cells (CHO-DG44(dfhr − / − )) which were dihydrofolate reductase deficient cells and high expression clones were selected by dot blotting. Further, gene amplification treatment was performed by adding load on cells in a medium containing 60 nM methotrexate (Mtx). Cells stably expressing rat-canine chimeric anti-PD-L1 antibody c4G12 (clone name: 4.3F1) after gene amplification were transferred to Mtx-free Opti-CHO medium and cultured under shaking for 14 days (125 rpm, 37° C., 5% CO 2 ). Cell survival rate was calculated by trypan blue staining ( FIG. 5 ). Chimeric antibody production in the culture supernatant was measured by ELISA ( FIG. 5 ). The culture supernatant at day 14 was centrifuged at 10,000 g for 10 min to remove cells, then passed through a 0.22 μm filter before the process proceeded to purification steps for the antibody. 
     It should be noted that by exchanging the medium with Dynamis medium and doing appropriate feeding, antibody production was improved about two-fold compared to the conventional production (data not shown). 
     2.9 Purification of Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 
     The culture supernatant provided as described above was purified with Ab Capcher Extra (ProteNova). An open column method was used for binding to resin; PBS pH 7.4 was used as equilibration buffer and wash buffer. As elution buffer, IgG Elution Buffer (Thermo Scientific) was used. As neutralization buffer, 1 M Tris was used. The purified antibody was concentrated and buffer-exchanged with PBS by ultrafiltration using Amicon Ultra-15 (50 kDa, Millipore). The resultant antibody was passed through a 0.22 μm filter for use in respective experiments. 
     2.10 Confirmation of Purification of Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 ( FIG. 6 ) 
     In order to confirm the purity of the purified antibody, antibody proteins were detected by SDS-PAGE and CBB staining. Using SuperSep Ace 5-20% (Wako) gradient gel, rat anti-bovine PD-L1 monoclonal antibody 4G12 and rat-canine chimeric anti-PD-L1 antibody c4G12 were electrophoresed under reducing conditions and non-reducing conditions. Bands were stained with Quick-CBB kit (Wako) and decolored in distilled water. Bands were observed at positions of molecular weights corresponding to antibodies. No bands of contaminant proteins were recognized visually. 
     2.11 Measurement of Binding Avidities to cPD-L1-His of Rat Anti-Bovine PD-L1 
     Monoclonal Antibody 4G12 and Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 In order to amplify the extracellular region of canine PD-L1 estimated from its putative amino acid sequence, primers were designed. Briefly, a primer having an NheI recognition sequence added on the 5′ side (cPD-L1-His F) and a primer having an EcoRV recognition sequence and 6×His tag sequence added on the 5′ side (cPD-L1-His R) were designed. PCR was performed using a synthesized beagle PBMC-derived cDNA as a template. The PCR products were digested with NheI (Takara) and EcoRV (Takara) and purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics). The thus purified DNA was cloned into pCXN2.1 vector (Niwa et al., 1991; kindly provided by Dr. T. Yokomizo, Juntendo University Graduate School of Medicine) treated with restriction enzymes in the same manner. The expression plasmids were purified with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pCXN2.1-cPD-L1-His. 
     
       
         
           
               
               
            
               
                   
                 Primer (cPD-L1-His F): 
               
               
                   
                 (SEQ ID NO: 131) 
               
               
                   
                 CGCGGCTAGCATGAGAATGTTTAGTGTCTT 
               
               
                   
                   
               
               
                   
                 Primer (cPD-L1-His R): 
               
               
                   
                 (SEQ ID NO: 132) 
               
               
                   
                 CGCGGATATCTTAATGGTGATGGTGATGGT 
               
               
                   
                   
               
               
                   
                 GAGTCCTCTCACTTGCTGG  
               
            
           
         
       
     
     The expression vector was transfected into Expi293F cells (Life Technologies) to obtain a culture supernatant containing a recombinant protein. The recombinant protein produced was purified from the supernatant using TALON Metal Affinity Resin (Clontech), and the buffer was exchanged with PBS using Amicon Ultra-4 Ultracel-3 (Merck Millipore). The thus obtained recombinant protein was stored at 4° C. until use in experiments (cPD-L1-His). The protein concentration was measured with Pierce BCA Protein Assay Kit (Thermo Fisher Scientific) for use in subsequent experiments. 
     Using a biomolecular interaction analyzer (Biacore X100), the binding avidities to cPD-L1-His of rat anti-bovine PD-L1 monoclonal antibody 4G12 and rat-canine chimeric anti-PD-L1 antibody c4G12 were assessed. Briefly, anti-histidine antibody was fixed on CM5 censor chip, followed by capturing of cPD-L1-His. Subsequently, monoclonal antibodies were added as analyte to observe specific binding. Both antibodies exhibited specific binding and their avidities were almost comparable (Table 1). Further, the binding avidities of canine PD-1-Ig and CD80-Ig to cPD-L1-His were measured in the same manner and found to be clearly lower than that of rat-canine chimeric anti-PD-L1 antibody c4G12 (Table 1). 
                     TABLE 1                  Binding Avidity of Each Antibody and Recombinant       Protein to Canine PD-L1-His                                         ka (×10 6 /Ms)   kd (×10 −3 /s)   KD (nM)                                                         4G12   2.42 ± 0.10   4.54 ± 0.19   1.88 ± 0.06               c4G12   3.14 ± 0.19   7.19 ± 0.26   2.30 ± 0.07               cPD-1           25.4 ± 4.89               cCD80           24.3 ± 0.89                        
2.12 Inhibitory Activity of Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 Against Canine PD-1/PD-L1 Binding and CD80/PD-L1 Binding ( FIG. 7 )
 
     Using the canine PD-1-Ig, PD-L1-Ig and CD80-Ig (described above), anti-PD-L1 antibody was tested for its ability to inhibit canine PD-1/PD-L1 binding and CD80/PD-L1 binding. Briefly, canine PD-1-Ig or CD80-Ig was immobilized on flat-bottom 96-well plates. Canine PD-L1-Ig was reacted with various concentrations (0, 2.5, 5 and 10 μg/ml) of rat anti-bovine PD-L1 antibody 4G12 or rat-canine chimeric anti-PD-L1 antibody c4G12 according to the same procedures as described in 2.6 above, and the binding of canine PD-L1-Ig was assessed. No change in binding inhibition activity was observed due to the chimerization of antibody. 
     2.13. Canine Immune Cell Activating Effect of Rat-Canine Chimeric Anti-PD-L1 Antibody c4G12 ( FIG. 8 ) 
     Canine PBMCs were cultured under stimulation with a super-antigen Staphylococcal Enterotoxin B (SEB) for three days, and changes in cytokine production by addition of rat-canine chimeric anti-PD-L1 antibody c4G12 were measured by ELISA using Duoset ELISA canine IL-2 or IFN-γ (R&amp;D systems). Rat-canine chimeric anti-PD-L1 antibody c4G12 increased the production of IL-2 and IFN-γ from canine PBMCs. Further, nucleic acid analogue EdU was added to the culture medium at day 2 of the culture under SEB stimulation. Two hours later, uptake of EdU was measured by flow cytometry using Click-iT Plus EdU flow cytometry assay kit (Life Technologies). As a result, EdU uptake in canine CD4 +  and CD8 +  lymphocytes was enhanced by addition of rat-canine chimeric anti-PD-L1 antibody c4G12, indicating an elevated cell proliferation capacity. 
     2.14 Selection of Tumor-Affected Dogs to be Used in Canine Inoculation Test 
     Since the subject treatment is expected to manifest a higher efficacy when PD-L1 is being expressed in tumors, PD-L1 expression analysis at the tumor site of dogs was performed by immunohistochemical staining. Briefly, tumor tissue samples fixed with formaldehyde and embedded in paraffin were sliced into 4 μm thick sections with a microtome, attached to and dried on silane-coated slide glass (Matsunami Glass) and deparaffinized with xylene/alcohol. While the resultant sections were soaked in citrate buffer [citric acid (Wako Pure Chemical) 0.37 g, trisodium citrate dihydrate (Kishida Chemical) 2.4 g, distilled water 1000 ml], antigen retrieval treatment was performed for 10 min with microwave, followed by staining using a Nichirei automatic immuno-staining device. As pretreatment, sample sections were soaked in 0.3% hydrogen peroxide-containing methanol solution at room temperature for 15 min and washed with PBS. Then, anti-bovine PD-L1 monoclonal antibody was added and reaction was conducted at room temperature for 30 min. After washing with PBS, histofine simple stain MAX-PO (Rat) (Nichirei Bioscience) was added and reaction was carried at room temperature for 30 min, followed by coloring with 3,3′-diaminobenzidine tetrahydrochloride and observation with a light microscope. Dogs with oral melanoma or undifferentiated sarcoma in which tumor cells were PD-L1 positive were used in the following inoculation test (clinical trial). Anti-bovine PD-L1 monoclonal antibody was established from a rat anti-bovine PD-L1 monoclonal antibody producing hybridoma (Ikebuchi R. Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y. Murata S, Ohashi K. Immunology. 2014 August; 142(4):551-61). 
     2.15 Inoculation Test on Dogs 
     With respect to the rat-canine chimeric anti-PD-L1 antibody c4G12 to be inoculated into dogs in the clinical trial, the culture supernatant obtained by the procedures described in 2.8 above was purified by affinity chromatography using MabSelect SuRe LX (GE Healthcare) and then by hydroxyapatite chromatography using BioScale CHT20-1 prepacked column (Bio-Rad) in order to remove contaminants and polymeric proteins. Aggregate-containing fractions were further purified by anion exchange chromatography using HiScreen Q-Sepharose HP prepacked column (GE Healthcare). 
     (1) Safety Test: The established rat-canine chimeric anti-PD-L1 antibody c4G12 was administered intravenously into a dog (beagle, spayed female, 13-year-old, about 10 kg in body weight) at 2 mg/kg, every 2 weeks, 3 times in total. There was observed no anaphylaxis or other adverse effects that would cause any trouble in clinical trials. (2) Clinical Trial 1: The established rat-canine chimeric anti-PD-L1 antibody c4G12 was administered intravenously into a PD-L1 positive dog with relapsed oral melanoma ( FIG. 9A ) (miniature dachshund, male, 11-year-old, about 7.5 kg in body weight) at 2 mg/kg or 5 mg/kg, every 2 weeks. 22 times in total. At week 10 after the start of treatment, a remarkable reduction in tumor size was recognized. At week 34 after the start of treatment, a still further reduction was confirmed ( FIG. 10 ). During the observation period of 44 weeks, no metastases to lymph nodes or the lung were observed. When 30% or more reduction in the longest diameter of tumor compared to that at the baseline is defined as PR (partial response), the criterion of PR was satisfied at weeks 16-20 and at week 34 and thereafter ( FIG. 11 ).
 
(3) Clinical Trial 2: Rat-canine chimeric anti-PD-L1 antibody c4G12 was administered intravenously into a dog with undifferentiated sarcoma whose primary lesion was PD-L1 positive ( FIG. 9B ) and who had a plurality of metastatic lesions in muscles throughout the body (west highland white terrier, castrated male, 12-year-old, about 8 kg in body weight) at 5 mg/kg, every 2 weeks, 2 times in total. At week 3 from the start of treatment, a clear regression of tumor was recognized ( FIG. 12 ).
 
(4) Clinical Trial 3: Rat-canine chimeric anti-PD-L1 antibody c4G12 was administered intravenously into a dog with oral melanoma whose primary lesion had been removed by surgery (beagle, spayed female, I1-year-old, about 10 kg in body weight) at 2 mg/kg or 5 mg/kg, every 2 weeks, 9 times in total. At week 18 after the start of treatment, a plurality of pulmonary metastatic lesions disappeared ( FIG. 13 ),
 
(5) Clinical Trial 4: Rat-canine chimeric anti-PD-L1 antibody c4G12 was administered intravenously into 4 dogs with oral melanoma with pulmonary metastasis at 2 mg/kg or 5 mg/kg, every 2 weeks. Although no clear reduction in tumor size was observed during the observation period, the duration of the treated dogs&#39; survival after confirmation of pulmonary metastasis tended to be longer than that of a control group (antibody not administered, historical control group: n=15) ( FIG. 14 ). Therefore, the survival duration may have been extended by antibody administration.
 
     2.16 CDR Analysis of Anti-PD-L1 Antibody 
     The complementarity-determining regions (CDRs) of rat anti-bovine PD-L1 antibody 4G12 were determined using NCBI IGBLAST (http://www.ncbi.nlm.nih.gov/igblast/). The results are shown in  FIG. 15 . 
     [Example 2] Application of Anti-PD-L1 Antibody to Other Animal Species 
     1.1 Identification of Ovine, Porcine and Water Buffalo PD-L1 Genes 
     In order to determine the full-lengths of the coding sequences (CDSs) of ovine, porcine and water buffalo PD-L1 cDNAs, primers for amplifying the full lengths of CDSs from the nucleotide sequences of ovine, porcine and water buffalo PD-L1 genes (GenBank accession number; XM_004004362, NM_001025221 and XM_613366) were first designed (ovPD-L1 CDS F and R; poPD-L1 CDS F and R; buPD-L1 CDS F1, R1, F2 and R2), and then PCR was performed. For the resultant amplified products, nucleotide sequences were determined with a capillary sequencer according to conventional methods (Mingala C N, Konnai S, Ikebuchi R, Ohashi K. Comp. Immunol. Microbiol. Infect. Dis. 2011 January; 34(1):55-63; Water buffalo PD-L1 gene was identified in this article). 
     
       
         
           
               
               
            
               
                   
                 Primer (ovPD-L1 CDS F): 
               
               
                   
                 (SEQ ID NO: 109) 
               
               
                   
                 ATGAGGATATATAGTGTCTTAACAT 
               
               
                   
                   
               
               
                   
                 Primer (ovPD-L1 CDS R): 
               
               
                   
                 (SEQ ID NO: 110) 
               
               
                   
                 TTACGICTCCTCAAAATGTG 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1 CDS F): 
               
               
                   
                 (SEQ ID NO: 111) 
               
               
                   
                 ATGAGGAFAIGTAGTATCTTTACAT 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1 CDS R): 
               
               
                   
                 (SEQ ID NO: 112) 
               
               
                   
                 TTACGTCTCCTCAAATTGTGT 
               
               
                   
                   
               
               
                   
                 Primer (buPD-L1 CDS F1): 
               
               
                   
                 (SEQ ID NO: 113) 
               
               
                   
                 ATGAGGATATATAGTGTCTT 
               
               
                   
                   
               
               
                   
                 Primer (buPD-L1 CDS R1): 
               
               
                   
                 (SEQ ID NO: 114) 
               
               
                   
                 GCCACTCAGGACTTGGTGAT 
               
               
                   
                   
               
               
                   
                 Primer (buPD-L1 CDS F2): 
               
               
                   
                 (SEQ ID NO: 115) 
               
               
                   
                 GGGGGITTACTGTTGCTTGA 
               
               
                   
                   
               
               
                   
                 Primer (buPD-L1 CDS R2): 
               
               
                   
                 (SEQ ID NO: 116) 
               
               
                   
                 TTACGTCTCCTCAAATTGT 
               
            
           
         
       
     
     1.2 Construction of Ovine PD-1, Ovine PD-L1, Porcine PD-1 and Porcine PD-L1 Expressing COS-7 Cells 
     In order to prepare ovine PD-1, ovine PD-L1, porcine PD-1 and porcine PD-L1 expressing plasmids, PCR was performed using a synthesized ovine or porcine PBMC-derived cDNA as a template and primers designed by adding BglII and SmaI (ovine PD-1), HindIII and SmaI (porcine PD-1), or XhoI and SmaI (ovine and porcine PD-L1) recognition sites on the 5′ side (ovPD-1-EGFP F and R; ovPD-L1-EGFP F and R; poPD-1-EGFP F and R; or poPD-L1-EGFP F and R). The resultant PCR products were digested with BglII (Takara) and SmaI (Takara) (ovine PD-1), HindIII (Takara) and SmaI (Takara) (porcine PD-1), and XhoI (Takara) and SmaI (Takara) (ovine and porcine PD-L1), then purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics) and cloned into pEGFP-N2 vector (Clontech) treated with restriction enzymes in the same manner. Expression plasmids were extracted using FastGene Xpress Plasmid PLUS Kit (NIPPON Genetics) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared plasmid is designated as pEGFP-N2-ovPD-1, pEGFP-N2-ovPD-L1, pEGFP-N2-poPD-1 or pEGFP-N2-poPD-L1. 
     
       
         
           
               
               
            
               
                   
                 Primer (ovPD-1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 117) 
               
               
                   
                 GAAGATCTATGGGGACCCCGCGGGCGCCG 
               
               
                   
                   
               
               
                   
                 Primer (ovPD-1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 118) 
               
               
                   
                 GACCCGGGGAGGGGCCAGGAGCAGTGTCC 
               
               
                   
                   
               
               
                   
                 Primer (ovPD-L1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 119) 
               
               
                   
                 CCGCTCGAGATGAGGATATATAGTGTCT 
               
               
                   
                   
               
               
                   
                 Primer (ovPD-L1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 120) 
               
               
                   
                 ATCCCGGGCGTCTCCTCAAAATGTGTAG 
               
               
                   
                   
               
               
                   
                 Primer (poPD-1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 121) 
               
               
                   
                 ACTAAGCTrATGGGGACCCCGCGGG 
               
               
                   
                   
               
               
                   
                 Primer (poPD-1-EGFPR): 
               
               
                   
                 (SEQ ID NO: 122) 
               
               
                   
                 ACTCCCGGGGAGGGGCCAAGAGCAGT 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 123) 
               
               
                   
                 CCGCTCGAGATGAGGATATGTAGTATCTT 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 124) 
               
               
                   
                 ATCCCGGGCGTCTCCTCAAATTGTGTATC 
               
            
           
         
       
     
     COS-7 cells were subcultured at a density of 5×10 4  cells/cm 2  in 6-well plates, and then cultured overnight in RPMI 1640 medium containing 10% inactivated fetal bovine serum and 0.01% L-glutamine at 37° C. in the presence of 5% CO 2 . The pEGFP-N2-ovPD-1, pEGFP-N2-ovPD-L1, pEGFP-N2-poPD-1, pEGFP-N2-poPD-L1 or pEGFP-N2 (negative control) was introduced into COS-7 cells at 0.4 μg/cm 2  using Lipofectamine 2000 (Invitrogen). The cells were cultured for 48 hours (ovPD-1-EGFP expressing cell, ovPD-L1-EGFP expressing cell, poPD-1-EGFP expressing cell, and poPD-L1-EGFP expressing cell). In order to confirm the expression of ovine PD-1, ovine PD-L1, porcine PD-1 and porcine PD-L1 in the thus prepared expressing cells, intracellular localization of EGFP was visualized with an inverted confocal laser microscope LSM700 (ZEISS) or an all-in-one fluorescence microscope BZ-9000 (KEYENCE). 
     1.3 Construction of Recombinant Ovine PD-L1 and Porcine PD-L1 
     In order to amplify the extracellular regions of ovine PD-L1 or porcine PD-L1 estimated from their putative amino acid sequences, primers were designed. Briefly, primers having an NheI or EcoRV recognition sequence added on the 5′ side (ovPD-L1-Ig F and R, or poPD-L1-Ig F and R) were designed. PCR was performed using a synthesized ovine or porcine PBMC-derived cDNA as a template. The PCR products were digested with NheI (Takara) and EcoRV (Takara) and purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics). The thus purified DNAs were individually cloned into pCXN2.1-Rabbit IgG Fc vector (Niwa et al., 1991. Zettimeissl et al., 1990, kindly provided by Dr. T. Yokomizo, Juntendo University Graduate School of Medicine, and modified in the inventors&#39; laboratory) treated with restriction enzymes in the same manner. The expression plasmids were purified with FastGene Xpress Plasmid PLUS Kit (NIPPON Genetics) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmids are designated as pCXN2.1-ovPD-L1-Ig and pCXN2.1-poPD-L1-Ig, respectively. 
     
       
         
           
               
               
            
               
                   
                 Primer (ovPD-L1-Ig F): 
               
               
                   
                 (SEQ ID NO: 125) 
               
               
                   
                 GACGCTAGCATGAGGATATATAGTGTCT 
               
               
                   
                   
               
               
                   
                 Primer (ovPD-L1-Ig R): 
               
               
                   
                 (SEQ 1D NO: 126) 
               
               
                   
                 GCTCTGATATCCCTCGTTTTTGCTGGAT 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1-Ig F): 
               
               
                   
                 (SEQ ID NO: 127) 
               
               
                   
                 GACGCTAGCATGAGGATATGTAGTATCTT 
               
               
                   
                   
               
               
                   
                 Primer (poPD-L1-Ig R): 
               
               
                   
                 (SEQ ID NO: 128) 
               
               
                   
                 AGCTTGATATCCCTCTTTCTTGCTGGATC 
               
            
           
         
       
     
     Thirty micrograms of pCXN2.1-ovPD-L1-Ig or pCXN2.1-poPD-L1-Ig was introduced into 7.5×10 7  Expi293F cells (Life Technologies) using Expifectamin (Life Technologies). After 6-day shaking culture, a culture supernatant was collected. The culture supernatant contained an Fc fusion recombinant protein. The produced Fc recombinant protein was purified from the supernatant using Ab-Capcher Extra (ProteNova). After purification, the buffer was exchanged with PBS (pH 7.4) using PD-10 Desalting Column (GE Healthcare). The resultant recombinant protein was stored at −30° C. until use in Experiment (ovine PD-L1-Ig). Concentrations of purified ovine PD-L1-Ig and porcine PD-L1-Ig were measured with Rabbit IgG ELISA Quantitation Set (BETHYL). For each washing operation in ELISA, Auto Palte Washer BIO WASHER 50 (DS Pharma Biomedical) was used. Absorbance was measured with Microplate Reader MTP-650FA (Corona Electric). 
     1.4 Reactivity of Rat Anti-Bovine PD-L1 Antibody 4G12 with Ovine and Porcine PD-L1 
     It was confirmed by flow cytometry that rat anti-bovine PD-L1 monoclonal antibody cross-reacts with ovine and porcine PD-L1. Ovine or Porcine PD-L1-EGFP expressing COS-7 cells were blocked with 10% inactivated goat serum supplemented PBS at room temperature for 15 min and reacted with 10 μg/ml of rat anti-bovine PD-L1 antibody 4G12 at room temperature for 30 min. After washing, the cells were reacted with allophycocyanine-labeled anti-rat Ig goat antibody (Beckman Coulter) at room temperature for 30 min. For analysis, FACS Verse (BD Bioscience) was used. As a negative control antibody, rat IgG2a (κ) isotype control (BD Bioscience) was used. For every washing operation and dilution of antibodies, 1% bovine serum albumin supplemented PBS was used. 
     Experimental results are shown in  FIG. 16 . It was confirmed that rat anti-bovine PD-L1 antibody 4G12 binds to ovine and porcine PD-L1. 
     1.5 Reactivity of Rat Anti-Bovine PD-L1 Antibody 4G12 with Water Buffalo Leukocytes 
     Peripheral blood of water buffalo (Bubalus bubalis; Asian water buffalo) was hemolyzed with ACK buffer to isolate leukocytes. After blocking with 10% inactivated goat serum supplemented PBS at room temperature for 15 min, reaction with rat anti-bovine PD-L1 antibody 4G12, peridinin-chlorophyll-protein complex/cyanin 5.5-labeled anti-bovine CD14 antibody (mouse IgG1, CAM36A, VMRD) and anti-bovine CD11b antibody (mouse IgG2b, CC126, AbD Serotec) was conducted at room temperature for 30 min. After washing, reaction with allophycocyanine-labelled anti-rat Ig goat antibody (Beckman Coulter) and fluorescein isothiocyanate-labeled anti-mouse IgG2 goat antibody (Beckman Coulter) was conducted at room temperature for 30 min. For analysis, FACS Calibur (BD Biosciences) was used. As a negative control antibody, rat IgG2a (κ) isotype control (BD Biosciences) was used. For every washing operation and dilution of antibodies, 10% inactivated goat serum supplemented PBS was used. 
     Experimental results are shown in  FIG. 17 . Rat anti-bovine PD-L1 antibody 4G12 strongly bound to blood macrophages (CD14 +  CD11b +  cells) of water buffalo. On the other hand, rat anti-bovine PD-L1 antibody 4G12 weakly bound to lymphocytes (CD14 −  CD11b −  cells) of water buffalo. This difference in binding property is believed to reflect the expression levels of PD-L1 in macrophages and lymphocytes. 
     1.6 Inhibition Test on Ovine or Porcine PD-1/PD-L1 Binding with Rat Anti-Bovine PD-L1 Antibody 4G12 
     Using ovine PD-1-EGFP expressing COS-7 cells and ovine PD-L1-Ig recombinant protein, or porcine PD-1-EGFP expressing COS-7 cells and porcine PD-L1-Ig recombinant protein, inhibition of ovine or porcine PD-1/PD-L1 binding by rat anti-bovine PD-L1 antibody (4G12) was tested. Briefly, rat anti-bovine PD-L1 antibody 4G12 of various concentrations (0, 1, 5, 10, 20, 50 μg/ml) was reacted in advance with ovine PD-L1-Ig (final concentration 1 μg/ml) or porcine PD-L1-Ig (final concentration 5 μg/ml) at 37° C. for 30 min. Subsequently, the antibody 4G12 was reacted with 2×10 5  ovine PD-1-EGFP expressing COS-7 cells or porcine PD-1-EGFP expressing COS-7 cells at 37° C. for 30 min. After washing, ovine PD-L1-Ig or porcine PD-L1-Ig bound to cell surfaces was detected with Alexa Fluor 647-labeled anti-rabbit IgG (H+L) goat F(ab′)2 (Life Technologies). For analysis, FACS Verse (BD Biosciences) was used. As a negative control antibody, rat IgG2a (κ) isotype control (BD Biosciences) was used. Taking the proportion of PD-L1-Ig bound cells without antibody addition as 100%, the proportion of PD-L1-Ig bound cells at each antibody concentration was shown as relative value. 
     The results revealed that rat anti-bovine PD-L1 antibody 4G12 is capable of inhibiting ovine PD-1/PD-L1 and porcine PD-1/PD-L1 binding in a concentration dependent manner ( FIG. 18 ). 
     Example 31 
     1. Introduction 
     Programmed cell death 1 (PD-1), an immunoinhibitory receptor, and its ligand programmed cell death ligand 1 (PD-L1) are molecules identified by Prof. Tasuku Honjo et al., Kyoto University, as factors which inhibit excessive immune response and are deeply involved in immunotolerance. Recently, it has been elucidated that these molecules are also involved in immunosuppression in tumors. In the subject Example, for the purpose of establishing a novel therapy for bovine infections, the present inventors have prepared a chimeric antibody gene by linking the variable region gene of rat anti-bovine PD-L1 monoclonal antibody (4G12) capable of inhibiting the binding of bovine PD-1 and PD-L1 to the constant region gene of a bovine immunoglobulin (IgG1 with mutations having been introduced into the putative binding sites for Fcγ receptors in CH2 domain to inhibit ADCC activity; see  FIG. 19  for amino acid numbers and mutations: 250 E→P, 251 L→V, 252 P→A, 253 G→deletion. 347 A→S, 348 P→S; Ikebuchi R. Konnai S, Okagawa T. Yokoyama K, Nakajima C. Suzuki Y, Murata S. Ohashi K. Immunology 2014 August; 142(4):551-561). This chimeric antibody gene was introduced into Chinese hamster ovary cells (CHO cells). By culturing/proliferating the resultant cells, the present inventors have obtained a rat-bovine chimeric anti-bovine PD-L1 antibody (ch4G12) and confirmed its effect in vitro and in vivo. 
     2. Materials and Methods 
     Construction of Bovine PD-1 and PD-L1 Expressing Cells 
     The nucleotide sequences of the full length cDNAs of bovine PD-1 gene (GenBank accession number AB510901; Ikebuchi R, Konnai S. Sunden Y, Onuma M, Ohashi K. Microbiol. Immunol. 2010 May; 54(5):291-298) and bovine PD-L1 gene (GenBank accession number AB510902: Ikebuchi R, Konnai S. Shirai T. Sunden Y. Murata S, Onuma M, Ohashi K. Vet. Res. 2011 Sep. 26; 42:103) were determined. Based on the resultant genetic information, bovine PD-1 and bovine PD-L1 membrane expressing cells were prepared. First, for preparing bovine PD-1 or PD-L1 expressing plasmid, PCR was performed using a synthesized bovine PBMC-derived cDNA as a template and designed primers having NotI and HindIII (bovine PD-1) recognition sites and NheI and XhoI (bovine PD-L1) recognition sites on the 5′ side (boPD-1-myc F and R; boPD-L1-EGFP F and R). The PCR products were digested with NotI (Takara) and HindIII (Takara; bovine PD-1), NheI (Takara) and XhoI (Takara; bovine PD-L1), purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics) and cloned into pCMV-Tag1 vector (Agilent Technologies; bovine PD-1) or pEGFP-N2 vector (Clontech; bovine PD-L1) treated with restriction enzymes in the same manner. The resultant expression plasmid of interest was extracted with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pCMV-Tag1-boPD-1. 
     
       
         
           
               
               
            
               
                   
                 Primer (boPD-1-mvc F): 
               
               
                   
                 (SEQ ID NO: 133) 
               
               
                   
                 ATATGCGGCCGCATGGGGACCCCGCGGGCGCT 
               
               
                   
                   
               
               
                   
                 Primer (boPD-1-mvc R): 
               
               
                   
                 (SEQ ID NO: 134) 
               
               
                   
                 GCGCAAGCTTTCAGAGGGGCCAGGAGCAGT 
               
               
                   
                   
               
               
                   
                 Primer (boPD-L1-EGFP F): 
               
               
                   
                 (SEQ ID NO: 135) 
               
               
                   
                 CTAGCTAGCACCATGAGGATATATAGTGTCTTAAC 
               
               
                   
                   
               
               
                   
                 Primer (boPD-L1-EGFP R): 
               
               
                   
                 (SEQ ID NO: 136) 
               
               
                   
                 CAATCTCGAGTTACAGACAGAAGATGACTGC 
               
            
           
         
       
     
     Bovine PD-1 membrane expressing cells were prepared by the procedures described below. First, 2.5 μg of pCMV-Tag1-boPD-1 was introduced into 4×10 6  CHO-DG44 cells using Lipofectamine LTX (Invitrogen). Forty-eight hours later, the medium was exchanged with CD DG44 medium (Life Technologies) containing 800 μg/ml G418 (Enzo Life Science). 20 ml/L GlutaMAX supplement (Life Technologies), and 18 ml/L 10% Pluronic F-68 (Life Technologies), followed by selection. The resultant expression cells were reacted with rat anti-bovine PD-1 antibody 5D2 at room temperature. After washing, the cells were further reacted with anti-rat IgG microbeads-labeled antibody (Miltenyi Biotec) at room temperature. Cells expressing bovine PD-1 at high levels were isolated with Auto MACS (Miltenyi Biotec). Subsequently, re-isolation was performed in the same manner to obtain still higher purity. The resultant expression cells were subjected to cloning by limiting dilution to thereby obtain a CHO DG44 cell clone expressing bovine PD-1 at high level (bovine PD-1 expressing cells). 
     Bovine PD-L1 membrane expressing cells were prepared by the procedures described below. First, 2.5 μg of pEGFP-N2-boPD-L1 or pEGFP-N2 (negative control) was introduced into 4×10 6  CHO-DG44 cells using Lipofectamine LTX (Invitrogen). Forty-eight hours later, the medium was exchanged with CD DG44 medium (Life Technologies) containing G418 (Enzo Life Science) 800 μg/ml. GlutaMAX supplement (Life Technologies) 20 ml/L, and 10% Pluronic F-68 (Life Technologies) 18 ml/L, followed by selection and cloning by limiting dilution (bovine PD-L1 expressing cell clone). In order to confirm the expression of bovine PD-L1 in the thus prepared expressing cell clone, intracellular localization of EGFP was visualized with an inverted confocal laser microscope LSM700 (ZEISS). 
     Construction of Soluble Bovine PD-1 and PD-L1 Bovine PD-1-Ig expressing plasmid was constructed by the procedures described below. Briefly, the signal peptide and the extracellular region of bovine PD-1 (GenBank accession number AB510901) were linked to the Fe domain of the constant region of a known bovine IgG1 (GenBank accession number X62916) to prepare a gene sequence. After codons were optimized for CHO cells, gene synthesis was performed in such a manner that NotI recognition sequence, KOZAK sequence, bovine PD-1 signal peptide sequence, bovine PD-1 gene extracellular region sequence, bovine IgG1 Fc region sequence, and XbaI recognition sequence would be located in the gene in this order. It should be noted here that bovine IgG1 was mutated to inhibit ADCC activity; more specifically, mutations were introduced into the putative binding sites for Fcγ receptors of CH2 domain (sites of mutation: 185 E→P, 186 L→V, 187 P→A, 189 G→deletion, 281 A→S, 282 P→S; Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C. Suzuki Y, Murata S, Ohashi K. Immunology 2014 August; 142(4):551-561; the amino acid sequence of PD-1-Ig and the sites of mutation are disclosed in  FIG. 2  of this article). The synthesized gene strand was digested with NotI (Takara) and XbaI (Takara), purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics), and incorporated into the cloning site (NotI and XbaI restriction enzyme recognition sequences downstream of PCMV and between INRBG and PABGH) of expression vector pDN11 (kindly provided by Prof. S. Suzuki, Hokkaido University Research Center for Zoonosis Control) treated with restriction enzymes in the same manner, whereby bovine PD-1-Ig expressing vector was constructed. The expression plasmid was purified with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pDN11-boPD-1-Ig. 
     Bovine PD-L1-Ig expressing plasmid was constructed by the procedures described below. In order to amplify the signal peptide and the extracellular region of bovine PD-L1 (GenBank accession number AB510902), primers were designed that had NheI and EcoRV recognition sites added on the 5′ side (boPD-L1-Ig F and R). PCR was performed using a synthesized bovine PBMC-derived cDNA as a template. The PCR products were digested with NheI (Takara) and EcoRV (Takara), purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics) and cloned into pCXN2.1-Rabbit IgG1 Fc vector (Niwa et al.. 1991: Zettlmeissl et al., 1990; kindly provided by Dr. T. Yokomizo, Juntendo University Graduate School of Medicine, and modified in the inventors&#39; laboratory) treated with restriction enzymes in the same manner. The expression plasmid was purified with QIAGEN Plasmid Midi kit (Qiagen) or FastGene Xpress Plasmid PLUS Kit (NIPPON Genetics) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pCXN2.1-boPD-L1-Ig. 
     
       
         
           
               
               
            
               
                   
                 Primer (boPD-L1-Ig F): 
               
               
                   
                 (SEQ ID NO: 137) 
               
               
                   
                 GCTAGCATGAGGATATATAGTGTCTTAAC 
               
               
                   
                   
               
               
                   
                 Primer (boPD-L1-Ig R): 
               
               
                   
                 (SEQ ID NO: 138) 
               
               
                   
                 GATATCATTCCTCTTTTTTGCTGGAT 
               
            
           
         
       
     
     Soluble bovine PD-1-Ig expressing cells were prepared by the procedures described below. Briefly, 2.5 μg of pDN11-boPD-1-Ig was introduced into 4×10 6  CHO-DG44 cells using Lipofectamine LTX (Invitrogen). Forty-eight hours later, the medium was exchanged with OptiCHO AGT medium (Life Technologies) containing 800 μg/ml G418 (Enzo Life Science) and 20 ml/L GlutaMAX supplement (Life Technologies). After cultured for 3 weeks, the cells were subjected to selection. Briefly, the concentrations of the Fc fusion recombinant protein in the culture supernatants of the resultant cell clones were measured by ELISA using anti-bovine IgG F(c) rabbit polyclonal antibody (Rockland) to thereby select those cell clones that express the Fc fusion recombinant protein at high levels. The resultant highly expressing cell clone was transferred to a G418-free medium and cultured under shaking for 14 days, followed by collection of a culture supernatant. The culture supernatant containing the Fc fusion recombinants protein was ultrafiltered with Centricon Plus-70 (Millipore). Then, the Fc fusion recombinant protein was purified with Ab-Capcher Extra (ProteNova). After purification, the buffer was exchanged with phosphate-buffered physiological saline (PBS; pH 7.4) using PD-10 Desalting Column (GE Healthcare). The resultant protein was stored at −30° C. until use in experiments (bovine PD-1-Ig). The concentration of the purified bovine PD-1-Ig was measured by ELISA using IgG F(c) rabbit polyclonal antibody (Rockland). For each washing operation in ELISA, Auto Plate Washer BIO WASHER 50 (DS Pharma Biomedical) was used. Absorbance was measured with Microplate Reader MTP-650FA (Corona Electric). 
     Soluble bovine PD-L1-Ig expressing cells were prepared by the procedures described below. Briefly, 30 μg of pCXN2.1-boPD-L1-Ig was introduced into 7.5×10 7  Expi293F cells (Life Technologies) using Expifectamine (Life Technologies). After 7-day culture under shaking, the culture supernatant was collected. The recombinant protein was purified from the supernatant using Ab-Capcher Extra (ProteNova; bovine PD-L1-Ig). After purification, the buffer was exchanged with PBS (pH 7.4) using PD MiniTrap G-25 (GE Healthcare). The resultant protein was stored at −30° C. until use in experiments (bovine PD-L1-Ig). The concentration of the purified bovine PD-L1-Ig was measured using Rabbit IgG ELISA Quantitation Set (Bethyl). For each washing operation in ELISA, Auto Plate Washer BIO WASHER 50 (DS Pharma Biomedical) was used. Absorbance was measured with Microplate Reader MTP-650FA (Corona Electric). 
     Preparation of Rat Anti-Bovine PD-L1 Monoclonal Antibody Producing Cells 
     Rat was immunized in the footpad with bovine PD-L1-Ig (Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C. Suzuki Y, Murata S, Ohashi K. Immunology 2014 August; 142(4):551-561: bovine PD-L1-Ig was prepared by the method disclosed in this article and used for immunization). Hybridomas were established by the iliac lymph node method to thereby obtain rat anti-bovine PD-L1 monoclonal antibody producing hybridoma 4G12. With respect to the method of establishment of rat anti-bovine PD-L1 monoclonal antibody, details are disclosed in the following non-patent document (Ikebuchi R. Konnai S, Okagawa T. Yokoyama K, Nakajima C, Suzuki Y, Murata S. Ohashi K. Vet. Res. 2013 Jul. 22; 44:59; Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y, Murata S. Ohashi K. Immunology 2014 August; 142(4):551-561). 
     Preparation of Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody Expressing Vector 
     Rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 was established by fusing the antibody constant regions of bovine IgG1 and Igλ with rat anti-bovine PD-L1 antibody 4G12 being used as an antibody variable region. 
     First, the genes of heavy chain and light chain variable regions were identified from a hybridoma that would produce rat anti-bovine PD-L1 antibody 4G12. Subsequently, a gene sequence was prepared in which the heavy chain and the light chain variable regions of the antibody 4G12 were linked to known constant regions of bovine IgG1 (heavy chain; modified from GenBank Accession number X62916) and bovine Igλ (light chain; GenBank Accession number X62917), respectively, and codon optimization was carried out [rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12: SEQ ID NOS: 105 and 106 (amino acid sequences), SEQ ID NOS: 107 and 108 (nucleotide sequences after codon optimization)]. It should be noted that in order to suppress the ADCC activity of bovine IgG1, mutations were added to the putative binding sites of Fcγ receptors in CH2 domain (See  FIG. 19  for amino acid numbers and mutations: 250 E→P. 251 L→V, 252 P→A, 253 G→deletion, 347 A→S, 348 P→S; Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y. Murata S. Ohashi K. Immunology 2014 August; 142(4):551-561). Then, the gene was artificially synthesized in such a manner that NotI recognition sequence, KOZAK sequence, chimeric antibody light chain sequence, poly-A addition signal sequence (PABGH), promoter sequence (PCMV), SacI recognition sequence, intron sequence (INRBG), KOZAK sequence, chimeric antibody heavy chain sequence and XbaI recognition sequence would be located in this order. The synthesized gene strand was digested with NotI (Takara) and XbaI (Takara), purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics) and cloned into the cloning site (NotI and XbaI restriction enzyme recognition sequences downstream of PCMV and between INRBG and PABGH) of expression plasmid pDC6 (kindly provided by Prof. S. Suzuki, Hokkaido University Research Center for Zoonosis Control) treated with restriction enzymes in the same manner ( FIG. 20 ). The resultant plasmid was extracted with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pDC6-boPD-L1ch4G12. 
     Expression of Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody 
     The pDC6-boPD-L1ch4G12 was transfected into CHO-DG44 cells (CHO-DG44 (dfhr − / − )) which were a dihydrofolate reductase deficient cell. Forty-eight hours later, the medium was exchanged with OptiCHO AGT medium (Life Technologies) containing 20 ml/L GlutaMAX supplement (Life Technologies). After cultured for 3 weeks, the cells were subjected to selection and cloning by limiting dilution. Subsequently, the concentrations of the chimeric antibody in the culture supernatants were measured by dot blotting and ELISA using anti-bovine IgG F(c) rabbit polyclonal antibody (Rockland) to thereby select high expression clones. Further, the selected clones expressing rat-bovine chimeric anti-bovine PD-L1 antibody at high levels were subjected to gene amplification treatment by adding a load with 60 nM methotrexate (Mtx)-containing medium. The thus established cell clone stably expressing rat-bovine chimeric anti-bovine PD-L1 antibody was transferred into Mtx-free Opti-CHO AGT medium and cultured under shaking for 14 days (125 rpm, 37° C., 5% CO 2 ). Chimeric antibody production in the culture supernatant was measured by ELISA using anti-bovine IgG F(c) rabbit polyclonal antibody (Rockland). For each washing operation in ELISA, Auto Plate Washer BIO WASHER 50 (DS Pharma Biomedical) was used. Absorbance was measured with Microplate Reader MTP-650FA (Corona Electric). The culture supernatant at day 14 was centrifuged at 10,000 g for 10 min to remove cells, and the centrifugal supernatant was passed through a Steritop-GP 0.22 μm filter (Millipore) for sterilization and then stored at 4° C. until it was subjected to purification. 
     Purification of Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody 
     From the culture supernatant prepared as described above, each chimeric antibody was purified using Ab Capcher Extra (ProteNova). An open column method was used for binding to resin; PBS pH 7.4 was used as an equilibration buffer and a wash buffer. As an elution buffer, IgG Elution Buffer (Thermo Fisher Scientific) was used. As a neutralization buffer, 1M Tris (pH 9.0) was used. The purified antibody was subjected to buffer exchange with PBS (pH 7.4) using PD-10 Desalting Column (GE Healthcare) and concentrated using Amicon Ultra-15 (50 kDa, Millipore). The thus purified chimeric antibody was passed through a 0.22 μm syringe filter (Millipore) for sterilization and stored at 4° C. until use in experiments. 
     Confirmation of the Purity of Purified Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody (FIG.  21 ) 
     In order to confirm the purity of purified rat-bovine chimeric anti-bovine PD-L1 antibody, antibody proteins were detected by SDS-PAGE and CBB staining. Using 10% acrylamide gel, the purified rat-bovine chimeric antibody was electrophoresed under reducing conditions (reduction with 2-mercaptoethanol from Sigma-Aldrich) and non-reducing conditions. Bands were stained with Quick-CBB kit (Wako) and decolored in distilled water. The results are shown in  FIG. 21 . Bands were observed at predicted positions, that is, at 25 kDa and 50 kDa under reducing conditions and at 150 kDa under non-reducing conditions. 
     Binding Specificity of Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody (FIG.  22 ) 
     It was confirmed by flow cytometry that the rat-bovine chimeric anti-bovine PD-L1 antibody specifically binds to the bovine PD-L1 expressing cells (described above). First, rat anti-bovine PD-L1 antibody 4G12 or rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 was reacted with bovine PD-L1 expressing cells at room temperature for 30 min. After washing, APC-labeled anti-rat Ig goat antibody (Southern Biotech) or Alexa Fluor 647-labeled anti-bovine IgG (H+L) goat F(ab′)2 (Jackson ImmunoResearch) was reacted at room temperature for 30 min. As negative control antibody, rat IgG2a (κ) isotype control (BD Biosciences) or bovine IgG1 antibody (Bethyl) was used. After washing, each rat antibody or rat-bovine chimeric antibody bound to cell surfaces was detected by FACS Verse (BD Biosciences). For every washing operation and dilution of antibody, PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used. 
     The experimental results are shown in  FIG. 22 . It was revealed that rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 binds to bovine PD-L1 expressing cells in the same manner as rat anti-bovine PD-L1 antibody 4G12. 
     Inhibitory Activity of Rat-Bovine Chimeric Anti-PD-L1 Antibody Against Bovine PD-1/PD-L1 Binding 
     (1) Binding Inhibition Test on Bovine PD-L1 Expressing Cells and Soluble Bovine PD-1 (FIG.  23 ) 
     Using bovine PD-L1 expressing cells (described above) and bovine PD-1-Ig (described above), bovine PD-1/PD-L1 binding inhibition by anti-bovine PD-L1 antibody was tested. First, 2×10 5  bovine PD-L1 expressing cells were reacted with various concentrations (0, 0.32, 0.63, 1.25, 2.5, 5 or 10 μg/ml) of rat anti-bovine PD-L1 antibody 4G12 or rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 at room temperature for 30 min. As negative control antibody, rat IgG2a (κ) isotype control (BD Biosciences) or bovine IgG1 antibody (Bethyl) was used. After washing, bovine PD-1-Ig labeled with biotin using Lightning-Link Type A Biotin Labeling Kit (Innova Bioscience) was added to a final concentration of 2 μg/ml, followed by reaction for another 30 min at room temperature. Subsequently, after washing, bovine PD-1-Ig bound to cell surfaces was detected with APC-labeled streptavidin (BioLegend). For analysis, FACS Verse (BD Biosciences) was used. For every washing operation and dilution of antibody, PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used. Taking the proportion of PD-1-Ig bound cells without antibody addition as 100%, the proportion of PD-1-Ig bound cells at each antibody concentration was shown as relative value. 
     The experimental results are shown in  FIG. 23 . It was revealed that like rat anti-bovine PD-L1 antibody 4G12, rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 is capable of inhibiting bovine PD-1/PD-L1 binding in a concentration dependent manner. 
     (2) Binding Inhibition Test on Bovine PD-1 Expressing Cells and Soluble Bovine PD-L1 (FIG.  24 ) 
     Using bovine PD-1 expressing cells (described above) and bovine PD-L1-Ig (described above), bovine pD-1/PD-L1 binding inhibition by anti-bovine PD-L1 antibody was tested. First, rat anti-bovine PD-L1 antibody 4G12 or rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 at a final concentration of 0, 0.32, 0.63, 1.25, 2.5, 5 or 10 μg/ml and bovine PD-L1-Ig at a final concentration of 1 μg/ml were placed in 96-well plates, where they were reacted at room temperature for 30 min. The resultant mixture was reacted with 2×10 5  bovine PD-1 expressing cells at room temperature for 30 min. As negative control antibody, rat IgG2a (κ) isotype control (BD Biosciences) or bovine IgG1 antibody (Bethyl) was used. After washing, Alexa Fluor 647-labeled anti-rabbit IgG (H+L) goat F(ab′)2 (Life Technologies) was reacted at room temperature for 30 min to thereby detect bovine PD-L1-Ig bound to cell surfaces. For analysis, FACS Verse (BD Biosciences) was used. For every washing operation and dilution of antibody, PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used. Taking the proportion of PD-L1-Ig bound cells without antibody addition as 100%, the proportion of PD-L1-Ig bound cells at each antibody concentration was shown as relative value. 
     The experimental results are shown in  FIG. 24 . It was revealed that like rat anti-bovine PD-L1 antibody 4G12, rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 is capable of inhibiting bovine PD-1/PD-L1 binding in a concentration dependent manner. 
     Biological Activity Test Using Rat-Bovine Chimeric Anti-Bovine PD-L1 Antibody 
     (1) Effect on Cell Proliferation (FIG.  25 ) 
     In order to confirm that bovine PD-1/PD-L1 binding inhibition by rat-bovine chimeric anti-PD-L1 antibody activates lymphocytes, a biological activity test was performed using cell proliferation as an indicator. Briefly, bovine PBMCs isolated from peripheral blood of healthy cattle were suspended in PBS to give a concentration of 10×10 6  cells/ml, and reacted with carboxyfluorescein succinimidyl ester (CFSE) at room temperature for 20 min. After washing twice with RPMI 1640 medium (Sigma-Aldrich) containing 10°/o inactivated fetal bovine serum (Cell Culture Technologies), antibiotics (streptomycin 200 μg/ml, penicillin 200 U/ml) (Life Technologies) and 0.01% L-glutamine (Life Technologies), the PBMCs were reacted with anti-bovine CD3 mouse antibody (WSU Monoclonal Antibody Center) at 4° C. for 30 min. After washing, the PBMCs were reacted with anti-mouse IgG1 microbeads (Miltenyi Biotec) at 4° C. for 15 min, followed by isolation of CD3-positive T cells using autoMACS™ Pro(Miltenyi Biotec). To the isolated CD3-positive T cells, anti-bovine CD3 mouse antibody (WSU Monoclonal Antibody Center) and anti-bovine CD28 mouse antibody (Bio-Rad) were added. Then, the cells were co-cultured with bovine PD-L1 expressing cells (CD3-positive T cells: bovine PD-L1 expressing cells=10:1) in the presence or absence of 10 μg/ml of rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12. As a control for antibodies, serum-derived bovine IgG (Sigma-Aldrich) was used; as a control for PD-L1 expressing cells, EGFP expressing cells transfected with pEGFP-N2 were used. After a 6-day coculture, cells were harvested and reacted with anti-bovine CD4 mouse antibody and anti-bovine CD8 mouse antibody (Bio-Rad) at room temperature for 30 min. For the labeling of antibodies, Zenon Mouse IgG1 Labeling Kits (Life Technologies) or Lightning-Link Kit (Innova Biosciences) was used. For analysis, FACS Verse (BD Biosciences) was used. For washing operation after culturing and dilution of antibody, PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used. 
     The experimental results are shown in  FIG. 25 . Proliferation of CD4-positive and CD8-positive T cells was significantly suppressed by co-culture with bovine PD-L1 expressing cells. It was revealed that rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 inhibits this suppression in CD4-positive T cells. 
     (2) Effect on IFN-γ Production (FIG.  26 ) 
     In order to confirm that bovine PD-1/PD-L1 binding inhibition by rat-bovine chimeric anti-PD-L1 antibody activates lymphocytes, a biological activity test was performed using IFN-γ production as an indicator. Briefly, PBMCs isolated from peripheral blood of BLV-infected cattle were suspended in RPMI medium (Sigma-Aldrich) containing 10% inactivated fetal bovine serum (Cell Culture Technologies), antibiotics (streptomycin 200 μg/ml, penicillin 200 U/ml) (Life Technologies) and 0.01% L-glutamine (Life Technologies) to give a concentration of 4×10 6  cells/ml. To the PBMCs, 10 μg/ml of rat anti-bovine PD-L1 antibody 4G12 or rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12, and 2% BLV-infected fetal lamp kidney cell (FLK-BLV) culture supernatant were added; culturing was then performed at 37° C. under 5% CO 2  for 6 days. As control antibodies, serum-derived rat IgG (Sigma-Aldrich) and serum-derived bovine IgG (Sigma-Aldrich) were used. After a 6-day culture, a culture supernatant was collected, and IFN-γ production was measured with Bovine IFN-γ ELISA Kit (BETYL). For each washing operation in ELISA, Auto Plate Washer BIO WASHER 50 (DS Pharma Biomedical) was used. Absorbance was measured with Microplate Reader MTP-650FA (Corona Electric). 
     The experimental results are shown in  FIG. 26 . It was revealed that rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 increases bovine PBMCs&#39; IFN-γ response to BLV antigen in the same manner as rat anti-bovine PD-L1 antibody 4G12 (n=10). 
     CDR Analysis of Rat Anti-Bovine PD-L1 Antibody 
     The complementarity-determining regions (CDRs) of rat anti-bovine PD-L1 antibody 4G12 were determined using NCBI IGBLAST (http://www.ncbi.nlm.nih.gov/igblast/). The results are shown in  FIG. 19 . 
     Inoculation Test on Cattle 
     Established rat-bovine chimeric anti-bovine PD-L1 antibody ch4G12 (about 26W mg; 1 mg/kg) was intravenously administered into experimentally BLV-infected calf (Holstein, male, 7 months old, 267 kg). Blood samples were collected chronologically from the infected calf, followed by isolation of PBMCs by density gradient centrifugation. 
     (1) Cell Proliferation Response of T Cells to BLV Antigen (FIG.  27 ) 
     Bovine PBMCs were suspended in PBS and reacted with CFSE at room temperature for 20 min. After washing twice with RPMI 1640 medium (Sigma-Aldrich) containing 10% inactivated fetal bovine serum (Cell Culture Technologies), antibiotics (streptomycin 200 μg/ml, penicillin 200 U/ml) (Life Technologies) and 0.01% L-glutamine (Life Technologies), the cell concentration was adjusted to 4×10 6  cells/ml using the same medium. Culture supernatant of 2% BLV-infected fetal lamp kidney cells (FLK-BLV) was added to the PBMCs, which were then cultured at 37° C. under 5% CO 2  for 6 days. As a control, culture supernatant of 2% BLV-not-infected fetal lamp kidney cells (FLK) was used. After a 6-day culture, PBMCs were collected and reacted with anti-bovine CD4 mouse antibody, anti-bovine CD8 mouse antibody and anti-bovine IgM mouse antibody (Bio-Rad) at 4° C. for 20 min. For the labeling of antibodies, Zenon Mouse IgG1 Labeling Kits (Life Technologies) or Lightning-Link Kit (Innova Biosciences) was used. For analysis, FACS Verse (BD Biosciences) was used. For every washing operation and dilution of antibody. PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used. 
     The experimental results are shown in  FIG. 27 . As a result of antibody administration, BLV-specific cell proliferation response of CD4-positive T cells increased compared to the response before administration. 
     (2) Changes in the BLV Provirus Load (FIG.  28 ) 
     DNA was extracted from isolated bovine PBMCs using Wizard DNA Purification kit (Promega). The concentration of the extracted DNA was quantitatively determined, taking the absorbance (260 nm) measured with Nanodrop 8000 Spectrophotometer (Thermo Fisher Scientific) as a reference. In order to measure the BLV provirus load in PBMCs, real time PCR was performed using Cycleave PCR Reaction Mix SP (TaKaRa) and Probe/Primer/Positive control for bovine leukemia virus detection (TaKaRa). Light Cycler 480 System II (Roche Diagnosis) was used for measurement. 
     The experimental results are shown in  FIG. 28 . The BLV provirus load significantly decreased until the end of test period compared to the load before administration. 
     All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety. 
     INDUSTRIAL APPLICABILITY 
     The anti-PD-L1 antibody of the present invention is applicable to prevention and/or treatment of cancers and infections in animals. 
     
       
         
           
               
               
            
               
                 SEQUENCE LISTING FREE TEXT 
                   
               
               
                 &lt;SEQ ID NO: 1&gt; 
                   
               
               
                 SEQ ID NO: 1 shows the amino acid sequence of the light chain variable region (VL) of rat 
                   
               
               
                 anti-bovine PD-L1antibody. 
               
               
                 MESQTHVLISLLLSVSGTYGDIAITQSPSSVAVSVGETVTLSCKSSQSLLYSENQKDYL 
               
               
                   
               
               
                 GWYQQKPGQTPKPLIYWATNRHTGVPDRFTGSGSGTDFTLIISSVQAEDLADYYCGQ 
               
               
                   
               
               
                 YLVYPFTFGPGTKLELK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 2&gt; 
                   
               
               
                 SEQ ID NO: 2 shows the amino acid sequence of the heavy chain variable region (VH) of rat 
                   
               
               
                 anti-bovine PD-L1antibody. 
               
               
                 MGWSQIILFLVAAATCVHSQVQLQQSGAELVKPGSSVKISCKASGYTFTSNFMHWVK 
               
               
                   
               
               
                 QQPGNGLEWIGWIYPEYGNTKYNQKFDGKATLTADKSSSTAYMQLSSLTSEDSAVYF 
               
               
                   
               
               
                 CASEEAVISLVYWGQGTLVTVSS 
               
               
                   
               
               
                 &lt;SEQ ID NO: 3&gt; 
                   
               
               
                 SEQ ID NO: 3 shows the amino acid sequence of the light chain constant region (CL) of a 
                   
               
               
                 canine antibody. 
               
               
                 QPKASPSVTLFPPSSEELGANKATLVCLISDFYPSGVTVAWKASGSPVTQGVETTKPSK 
               
               
                   
               
               
                 QSNNKYAASSYLSLTPDKWKSHSSFSCLVTHEGSTVEKKVAPAECS 
               
               
                   
               
               
                 &lt;SEQ ID NO: 4&gt; 
                   
               
               
                 SEQ ID NO: 4 shows the amino acid sequence of the heavy chain constant region (CH) of a 
                   
               
               
                 canine antibody. 
               
               
                 ASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEPVTVSWNSGSLTSGVHTFPSVLQS 
               
               
                   
               
               
                 SGLYSLSSTVTVPSSRWPSETFTCNVVHPASNTKVDKPVPKESTCKCISPCPVPESLGG 
               
               
                   
               
               
                 PSVFIFPPKPKDILRITRTPEITCVVLDLGREDPEVQISWFVDGKEVHTAKTQPREQQFN 
               
               
                   
               
               
                 STYRVVSVLPIEHQDWLTGKEFKCRVNHIGLPSPIERTISKARGQAHQPSVYVLPPSPK 
               
               
                   
               
               
                 ELSSSDTVTLTCLIKDFFPPEIDVEWQSNGQPEPESKYHTTAPQLDEDGSYFLYSKLSV 
               
               
                   
               
               
                 DKSRWQQGDTFTCAVMIHEALQNHYTDLSLSHSPGK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 5&gt; 
                   
               
               
                 SEQ ID NO: 5 shows the nucleotide sequence of the VL of rat anti-bovine PD-L1 antibody. 
                   
               
               
                 ATGGAATCACAGACGCATGTCCTCATTTCCCTTCTGCTCTCGGTATCTGGTACCTAT 
               
               
                   
               
               
                 GGGGACATTGCGATAACCCAGTCTCCATCCTCTGTGGCTGTGTCAGTAGGAGAGA 
               
               
                   
               
               
                 CGGTCACTCTGAGCTGCAAGTCCAGTCAGAGTCTTTTATACAGTGAAAACCAAAA 
               
               
                   
               
               
                 GGACTATTTGGGCTCGGTACCAGCAGAAACCAGGGCAGACTCCTAAACCCCTTATCT 
               
               
                   
               
               
                 ACTGGGCAACCAACCGGCACACTGGGGTCCCTGATCGCTTCACAGGTAGTGGATC 
               
               
                   
               
               
                 CGGGACAGACTTCACTCTGATCATCAGCAGTGTGCAGGCTGAAGACCTGGCTGAT 
               
               
                   
               
               
                 TATTACTGTGGGCAGTACCTTGTCTATCCGTTCACGTTTGGACCTGGGACCAAGCT 
               
               
                   
               
               
                 GGAACTGAAA 
               
               
                   
               
               
                 The nucleotide sequence of SEQ ID NO: 5 after codon optimization is shown in &lt;SEQ ID 
               
               
                 NO: 15&gt;. 
               
               
                 ATGGAATCTCAAACTCATGTTTTGATTTCATTACTTCTGAGTGTTTCCGGAACCTAC 
               
               
                   
               
               
                 GGTGATATCGCTATCACTCAATCTCCCTCCTCTGITGCTGTGTCTGTGGGCGAAAC 
               
               
                   
               
               
                 CGTTACCCTGTCCTGCAAGTCCAGTCAGTCTCTTCTCTACTCCGAGAATCAAAAGG 
               
               
                   
               
               
                 ACTACCTGGGCTGGTACCAACAGAAGCCCGGCCAGACCCCAAAGCCACTGATATA 
               
               
                   
               
               
                 CTGGGCAACCAACAGGCACACCGGAGTGCCCGACAGGTTCACAGGCAGTGGATC 
               
               
                   
               
               
                 TGGCACCGACTTTACCTTGATCATTTCAAGCGTGCAGGCTGAAGATCTGGCCGACT 
               
               
                   
               
               
                 ACTACTGTGGTCAGTATCTGGTGTATCCTTTCACTTTCGGGCCAGGGACAAAATTG 
               
               
                   
               
               
                 GAATTGAAG 
               
               
                   
               
               
                 &lt;SEQ ID NO: 6&gt; 
                   
               
               
                 SEQ ID NO: 6 shows the nucleotide sequence of the VH of rat anti-bovine PD-L1 antibody. 
                   
               
               
                 ATGGGATGGAGCCAGATCATCCTCTTTCTGGTGGCAGCAGCTACATGTGTTCACTC 
               
               
                   
               
               
                 CCAGGTACAGCTGCAGCAATCTGGGGCTGAATTAGTGAAGCCTGGGTCCTCAGTG 
               
               
                   
               
               
                 AAAATTTCCTGCAAGGCTTCTGGCTACACCTTCACCAGTAACTTTATGCACTGGGT 
               
               
                   
               
               
                 AAAGCAGCAGCCTGGAAATGGCCTTGAGTGGATTGGGTGGATTTATCCTGAATATG 
               
               
                   
               
               
                 GTAATACTAAGTACAATCAAAAGTTCGATGGGAAGGCAACACTCACTGCAGACAA 
               
               
                   
               
               
                 ATCCTCCAGCACAGCCTATATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCAG 
               
               
                   
               
               
                 TCTATTTCTGTGCAAGTGAGGAGGCAGTTATATCCCTTGTTTACTGGGGCCAAGGC 
               
               
                   
               
               
                 ACTCTGGTCACTGTCTCTTCA 
               
               
                   
               
               
                 The nucleotide sequence of SEQ ID NO: 6 after codon optimization is shown in &lt;SEQ ID 
               
               
                 NO: 16&gt;. 
               
               
                 ATGGGTTGGTCTCAAATTATCTTGTTTTTGGTTGCTGCAGCCACTTGTGTTCATTCT 
               
               
                   
               
               
                 CAGGTGCAGCTGCAACAAAGCGGCGCAGAACTGGTGAAACCTGGCAGCAGCGTG 
               
               
                   
               
               
                 AAAATATCTTGTAAGGCCAGCGGATATACTTTCACCTCCAATTTCATGCATTGGGTC 
               
               
                   
               
               
                 AAACAGCAGCCCGGCAACGGACTCGAGTGGATCGGCTGGATCTACCCCGAGTATG 
               
               
                   
               
               
                 GCAACACAAAATATAACCAAAAATTTGATGAAAGGCTACCCTGACTGCCGATAA 
               
               
                   
               
               
                 GTCCTCCAGCACCGCATACATGCAACTCTCCTCCCTGACCTCCGAGGATAGCGCTG 
               
               
                   
               
               
                 TCTACTTCTGTGCTTCCGAAGAGGCTGTCATATCCTTGGTCTATTGGGGCCAAGGA 
               
               
                   
               
               
                 ACTCTGGTGACCGTCTCATCT 
               
               
                   
               
               
                 &lt;SEQ ID NO: 7&gt; 
                   
               
               
                 SEQ ID NO: 7 shows the nucleotide sequence of the CL of a canine antibody. 
                   
               
               
                 CAGCCCAAGGCCTCCCCCTCGGTCACACTCTTCCCGCCCTCCTCTGAGGAGCTCG 
               
               
                   
               
               
                 GCGCCAACAAGGCCACCCTGGTGTGCCTCATCAGCGACTTCTACCCCAGCGGCGT 
               
               
                   
               
               
                 GACGGTGGCCTGGAAGGCAAGCGGCAGCCCCGTCACCCAGGGCGTGGAGACCAC 
               
               
                   
               
               
                 CAAGCCCTCCAAGCAGAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCT 
               
               
                   
               
               
                 GACGCCTGACAAGTGGAAATCTCACAGCAGCTTCAGCTGCCTGGTCACGCACGA 
               
               
                   
               
               
                 GGGGAGCACCGTGGAGAAGAAGGTGGCCCCCGCAGAGTGCTCTTAG 
               
               
                   
               
               
                 The nucleotide sequence of SEQ ID NO: 7 after codon optimization is shown in &lt;SEQ ID 
               
               
                 NO: 17&gt;. 
               
               
                 CAGCCCAAAGCCTCTCCCAGCGTCACCCTCTTCCCACCTTCCAGTGAGGAGCTGG 
               
               
                   
               
               
                 GGGCAAACAAAGCCACTTTGGTGTGTCTCATCTCCGATTTTTACCCCTCCGGGGTC 
               
               
                   
               
               
                 ACAGTCGCATGGAAGGCCTCCGGATCCCCTGTGACACAGGGAGTGGAGACAACA 
               
               
                   
               
               
                 AAACCTAGCAAGCAGAGTAACAAATAAGTATGCCGCCTCAAGCTATCAGCCTTAC 
               
               
                   
               
               
                 TCCTGATAAGTGGAAGTCACATAGCAGTTTTAGTTGCCTCGTAACACATGAGGGTT 
               
               
                   
               
               
                 CAACTGTGGAGAAAAAAGTAGCTCCAGCTGAGTGCTCATGA 
               
               
                   
               
               
                 &lt;SEQ ID NO: 8&gt; 
                   
               
               
                 SEQ ID NO: 8 is the nucleotide sequence of the CH of a canine antibody. 
                   
               
               
                 GCCTCCACCACGGCCCCCTCGGTTTTCCeCACTGGCCCCCAGCTGCGGGTCCACTT 
               
               
                   
               
               
                 CCGGCTCCACGGTGGCCCTGGCCTGCCTGGTGTCAGGCTACTTCCCCGAGCCTGT 
               
               
                   
               
               
                 AACTGTGTCCTGGAATTCCGGCTCCTTGACCAGCGGTGTGCACACCTTCCCGTCC 
               
               
                   
               
               
                 GTCCTGCAGTCCTCAGGGCTCTACTCCCTCAGCAGCACGGTGACAGTGCCCTCCA 
               
               
                   
               
               
                 GCAGGTGGCCCAGCGAGACCTTCACCTGCAACGTGGTCCACCCGGCCAGCAACA 
               
               
                   
               
               
                 CTAAAGTAGACAAGCCAGTGCCCAAAGAGTCCACCTGCAAGTGTATATCCCCATG 
               
               
                   
               
               
                 CCCAGTCCCTGAATCACTGGGAGGGCCTTCGGTCTTCATCTTTCCCCCGAAACCCA 
               
               
                   
               
               
                 AGGACATCCTCAGGATTACCCGAACACCCGAGATCACCTGTGTGGTGTTAGATCTG 
               
               
                   
               
               
                 GGCCGTGAGGACCCTGAGGTGCAGATCAGCTGGTTCGTGGATGGTAAGGAGGTG 
               
               
                   
               
               
                 CACACAGCCAAGACGCAGCCTCGTGAGCAGCAGTTCAACAGCACCTACCGTGTG 
               
               
                   
               
               
                 GTCAGCGTCCTCCCCATTGAGCACCAGGACTGGCTCACCGGAAAGGAGTTCAAGT 
               
               
                   
               
               
                 GCAGAGTCAACCACATAGGCCTCCCGTCCCCCATCGAGAGGACTATCTCCAAAGC 
               
               
                   
               
               
                 CAGAGGGCAAGCCCATCAGCCCAGTGTGTATGTCCTGCCACCATCCCCAAAGGAG 
               
               
                   
               
               
                 TTGTCATCCAGTGACACGGTCACCCTGACCTGCCTGATCAAAGACTTCTTCCCACC 
               
               
                   
               
               
                 TGAGATTGATGTGGAGTGGCAGAGCAATGGACAGCCGGAGCCCGAGAGCAAGTA 
               
               
                   
               
               
                 CCACACGACTGCGCCCCAGCTGGACGAGGACGGGTCCTACTTCCTGTACAGCAAG 
               
               
                   
               
               
                 CTCTCTGTGGACAAGAGCCGCTGGCAGCAGGGAGACACCTTCACATGTGCGGTGA 
               
               
                   
               
               
                 TGCATGAAGCTCTACAGAACCACTACACAGATCTATCCTCTCCCATTCTCCGGGT 
               
               
                   
               
               
                 AAATGA 
               
               
                   
               
               
                 The nucleotide sequence of SEQ ID NO: 8 after codon optimization is shown in &lt;SEQ ID 
               
               
                 NO: 18&gt;. 
               
               
                 GCTAGCACAACCGCTCCCTCCGTTTTTCCCCTCGCCCCATCCTGCGGGTCAACCAG 
               
               
                   
               
               
                 CGGATCCACCGTCGCTCTGGCTTGTCTGGTGTCAGGATACTTCCCCGAGCCTGTCA 
               
               
                   
               
               
                 CCGTTTCTTGGAATAGCGGCAGCCTTACTTCCGGCGTGCATACCTTCCCTAGCGTG 
               
               
                   
               
               
                 CTTCAGTCCTCCGGTCTGTATTCCCTCAGCTCCACCGTAACTGTCCCAAGCTCAAG 
               
               
                   
               
               
                 GTGGCCCTCTGAGACATTTACCTGCAATGTGGTCCATCCTGCTTCAAATACCAAAG 
               
               
                   
               
               
                 TGGACAAGCCCGTCCCAAAAGAGTCTACCTGCAAATGTATCAGTCCTTGTCCCGT 
               
               
                   
               
               
                 GCCCGAGTCTCTGGGCGGACCCTCAGTCTTTATCTTCCCACCCAAGCCAAAGGAC 
               
               
                   
               
               
                 ATATTGCGCATTACACGGACACCCGAAATCACCTGTGTTGTGTTGGATCTCGGCCG 
               
               
                   
               
               
                 GGAAGATCCTGAGGTGCAGATTAGTTGGTTTGTTGATGGCAAGGAGGTGCACACA 
               
               
                   
               
               
                 GCAAAAACACAGCCCAGAGAACAGCAGTTCAACAGTACTTATAGAGTAGTGAGT 
               
               
                   
               
               
                 GTGTTGCCTATAGAGCATCAGGACTGGCTGACAGGCAAAGAATTCAAATGTAGGG 
               
               
                   
               
               
                 TTAACCACATTGGCCTCCCTAGTCCAATCGAGAGGACAATCTCTAAAGCCCGAGG 
               
               
                   
               
               
                 CCAGGCTCATCAGCCTTCTGTGTACGTTCTGCCTCCTAGTCCTAAGGAACTGTCTT 
               
               
                   
               
               
                 CTTCAGACACAGTAACACTCACTTGCCTGATTAAGGACTTTTTTCCTCCAGAGATT 
               
               
                   
               
               
                 GATGTGGAATGGCAGTCTAACGGGCAGCCAGAGCCAGAATCTAAGTACCACACTA 
               
               
                   
               
               
                 CTGCACCACAGCTGGATGAGGATGGGTCTTACTTCCTGTACAGTAAGCTGAGTGT 
               
               
                   
               
               
                 GGACAAGTCTCGATGGCAGCAGGGGGATACTTTTACTTGCGCAGTAATGCACGAA 
               
               
                   
               
               
                 GCATTGCAGAACCACTACACTGACCTGTCACTTAGTCACTCACCAGGGAAGTAA 
               
               
                   
               
               
                 &lt;SEQ ID NO: 9&gt; 
                   
               
               
                 SEQ ID NO: 9 shows the amino acid sequence of a chimeric light chain consisting of the VL 
                   
               
               
                 of rat anti-bovine Pd-L1 antibody and the CL of a canine antibody. 
               
               
                 MESQTHVLISLLLSVSGTYGDIAITQSPSSVAVSVGETVTLSCKSSQSLLYSENQKDYL 
               
               
                   
               
               
                 GWYQQKPGQTPKPLIYWATNRHTGVPDRFTGSGSGTDFTLIISSVQAEDLADYYCGQ 
               
               
                   
               
               
                 YLVYPFTFGPGTKLELKQPKASPSVTLFPPSSEELGANKATLVCLISDFYPSGVTVAWK 
               
               
                   
               
               
                 ASGSPVTQGVETTKPSKQSNNKYAASSYLSLTPDKWKSHSSFSCLVTHEGSTVEKKV 
               
               
                   
               
               
                 APAECS 
               
               
                   
               
               
                 &lt;SEQ ID NO: 10&gt; 
                   
               
               
                 SEQ ID NO: 10 shows the amino acid sequence of a chimeric heavy chain consisting of the 
                   
               
               
                 VH of rat anti-bovine PD-L1 antibody and the CH of a canine antibody. 
               
               
                 MGWSQIILFLVAAAFCVHSQVQLQQSGAELVKPGSSVKISCKASGYTFTSNFMHWVK 
               
               
                   
               
               
                 QQPGNGLEWIGWIYPEYGNTKYNQKFDGKATLTADKSSSTAYMQLSSLTSEDSAVYF 
               
               
                   
               
               
                 CASEEAVISLVYWGQGTLVTVSSASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEP 
               
               
                   
               
               
                 VTVSWNSGSLTSGVHTFPSVLQSSGLYSLSSTVTVPSSRWPSETFTCNVVHPASNTKV 
               
               
                   
               
               
                 DKPVPKESTCKCISPCPVPESLGGPSVFIFPPKPKDILRITRTPEITCVVLDLGREDPEVQ 
               
               
                   
               
               
                 ISWFVDGKEVHTAKTQPREQQFNSTYRVVSVLPIEHQDWLTGKEFKCRVNHIGLPSPI 
               
               
                   
               
               
                 ERTISKARGQAHQPSVYVLPPSPKELSSSDTVTLTCLIKDFFPPEIDVEWQSNGQPEPES 
               
               
                   
               
               
                 KYHTTAPQLDEDGSYFLYSKLSVDKSRWQQGDTFTCAVMHEALQNHYTDLSLSHSP 
               
               
                   
               
               
                 GK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 19&gt; 
                   
               
               
                 This sequence shows the nucleotide sequence (after codon optimization) of a chimeric light 
                   
               
               
                 chain consisting of the VL of rat anti-bovine PD-L1 antibody and the CL of a canine 
               
               
                 antibody. 
               
               
                 ATGGAATCTCAAACTCATGTTTTGATTTCATTACTTCTGAGTGTTTCCGGAACCTAC 
               
               
                   
               
               
                 GGTGATATCGCTATCACTCAATCTCCCTCCTCTGTTGCTGTGTCTGTGGGCGAAAC 
               
               
                   
               
               
                 CGTTACCCTGTCCTGCAAGTCCAGTCAGTCTCTTCTCTACTCCGAGAATCAAAAGG 
               
               
                   
               
               
                 ACTACCTGGGCTGGTACCAACAGAAGCCCGGCCAGACCCCAAAGCCACTGATATA 
               
               
                   
               
               
                 CTGGGCAACCAACAGGCACACCGGAGTGCCCGACAGGTTCACAGGCAGTGGATC 
               
               
                   
               
               
                 TGGCACCGACTTTACCTTGATCATTTCAAGCGTGCAGGCTGAAGATCTGGCCGACT 
               
               
                   
               
               
                 ACTACTGTGGTCAGTATCTGGTGTATCCTTTCACTTTCGGGCCAGGGACAAAATTG 
               
               
                   
               
               
                 GAATTGAAGCAGCCCAAAGCCTCTCCCAGCGTCACCCTCTTCCCACCTTCGAGTG 
               
               
                   
               
               
                 AGGAGCTGGGGGCAAACAAAGCCACTTTGGTGTGTCTCATCTCCGATTTTTACCC 
               
               
                   
               
               
                 CTCCGGGGTCACAGTCGCATGGAAGGCCTCCGGATCCCCTGTGACACAGGGAGTG 
               
               
                   
               
               
                 GAGACAACAAAACCTAGCAAGCAGAGTAACAATAAGTATGCCGCCTCAAGCTATC 
               
               
                   
               
               
                 TCAGCCTTACTCCTGATAAGTGGAAGTCACATAGCAGTTTTAGTTGCCTCGTAACA 
               
               
                   
               
               
                 CATGAGGGTTCAACTGTGGAGAAAAAAGTAGCTCCAGCTGAGTGCTCATGA 
               
               
                   
               
               
                 &lt;SEQ ID NO: 20&gt; 
                   
               
               
                 SEQ ID NO: 20 shows the nucleotide sequence (after codon optimization) of a chimeric 
                   
               
               
                 heavy chain consisting of the VH of rat anti-bovine PD-L1 antibody and the CH of a canine 
               
               
                 antibody. 
               
               
                 ATGGGTTGGTCTCAAATTATCTTGTTTTTGGTTGCTGCAGCCACTTGTGTTCATTCT 
               
               
                   
               
               
                 CAGGTGCAGCTGCAACAAAGCGGCGCAGAACTGGTGAAACCTGGCAGCAGCGTG 
               
               
                   
               
               
                 AAAATATCTTGTAAGGCCAGCGGATATACTTTCACCTCCAATTTCATGCATTGGGTC 
               
               
                   
               
               
                 AAACAGCAGCCCGGCAACGGACTCGAGTGGATCGGCTGGATCTACCCCGAGTATG 
               
               
                   
               
               
                 GCAACACAAAATATAACCAAAAATTTGATGGAAAGGCTACCCTGACTGCCGATAA 
               
               
                   
               
               
                 GTCCTCCAGCACCGCATACATGCAACTCTCCTCCCTGACCTCCGAGGATAGCGCTG 
               
               
                   
               
               
                 TCTACTTCTGTGCTTCCGAAGAGGCTGTCATATCCTTGGTCTATTGGGGCCAAGGA 
               
               
                   
               
               
                 ACTCTGGTGACCGTCTCATCTGCTAGCACAACCGCTCCCTCCGTTTTTCCCCTCGC 
               
               
                   
               
               
                 CCCATCCTGCGGGTCAACCAGCGGATCCACCGTCGCTCTGGCTTGTCTGGTGTCA 
               
               
                   
               
               
                 GGATACTTCCCCGAGCCTGTCACCGTTTCTTGGAATAGCGGCAGCCTTACTTCCGG 
               
               
                   
               
               
                 CGTGCATACCTTCCCTAGCGTGCTTCAGTCCTCCGGTCTGTATTCCCTCAGCTCCAC 
               
               
                   
               
               
                 CGTAACTGTCCCAAGCTCAAGGTGGCCCTCTGAGACATTTACCTGCAATGTGGTCC 
               
               
                   
               
               
                 ATCCTGCTTCAAATACCAAAGTGGACAAGCCCGTCCCAAAAGAGTCTACCTGCAA 
               
               
                   
               
               
                 ATGTATCAGTCCTTGTCCCGTGCCCGAGTCTCTGGGCGGACCCTCAGTCTTTATCTT 
               
               
                   
               
               
                 CCCACCCAAGCCAAAGGACATATTGCGCATTACACGGACACCCGAAATCACCTGT 
               
               
                   
               
               
                 GTTGTGTTGGATCTCGGCCGGGAAGATCCTGAGGTGCAGATTAGTTGGTTTGTTGA 
               
               
                   
               
               
                 TGGCAAGGAGGTGCACACAGCAAAAACACAGCCCAGAGAACAGCAGTTCAACA 
               
               
                   
               
               
                 GTACTTATAGAGTAGTGAGTGTGTTGCCTATAGAGCATCAGGACTGGCTGACAGGC 
               
               
                   
               
               
                 AAAGAATTCAAATGTAGGGTTAACCACATTGGCCTCCCTAGTCCAATCGAGAGGA 
               
               
                   
               
               
                 CAATCTCTAAAGCCCGAGGCCAGGCTCATCAGCCTTCTGTGTACGTTCTGCCTCCT 
               
               
                   
               
               
                 AGTCCTAAGGAACTGTCTTCTTCAGACACAGTAACACTCACTTGCCTGATTAAGG 
               
               
                   
               
               
                 ACTTTTTTCCTCCAGAGATTGATGTGGAATGGCAGTCTAACGGGCAGCCAGAGCC 
               
               
                   
               
               
                 AGAATCTAAGTACCACACTACTGCACCACAGCTGGATGAGGATGGGTCTTACTTCC 
               
               
                   
               
               
                 TGTACAGTAAGCTGAGTGTGGACAAGTCTCGATGGCAGCAGGGGGATACTTTTAC 
               
               
                   
               
               
                 TTGCGCAGTAATGCACGAAGCATTGCAGAACCACTACACTGACCTGTCACTTAGT 
               
               
                   
               
               
                 CACTCACCAGGGAAGTAA 
               
               
                   
               
               
                 &lt;SEQ ID NO: 11&gt; 
                   
               
               
                 SEQ ID NO: 11 shows the amino acid sequence of the CL of a human antibody. 
                   
               
               
                 TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE 
               
               
                   
               
               
                 QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 
               
               
                   
               
               
                 &lt;SEQ ID NO: 12&gt; 
                   
               
               
                 SEQ ID NO: 12 shows the amino acid sequence of the CH (CH1—CH3) of a human antibody 
                   
               
               
                 (IgG4 variant 1). 
               
               
                 STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS 
               
               
                   
               
               
                 GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEELGGP 
               
               
                   
               
               
                 SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE 
               
               
                   
               
               
                 QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTL 
               
               
                   
               
               
                 PPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSR 
               
               
                   
               
               
                 LTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 13&gt; 
                   
               
               
                 SEQ ID NO: 13 shows the nucleotide sequence of the CL of a human antibody. 
                   
               
               
                 ACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATC 
               
               
                   
               
               
                 TGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAA 
               
               
                   
               
               
                 GTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCA 
               
               
                   
               
               
                 CAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGA 
               
               
                   
               
               
                 GCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGG 
               
               
                   
               
               
                 GCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG 
               
               
                   
               
               
                 &lt;SEQ ID NO: 14&gt; 
                   
               
               
                 SEQ ID NO: 14 shows the nucleotide sequence of the CH (CH1—CH3) of a human antibody 
                   
               
               
                 (IgG4 variant 1). 
               
               
                 TCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCG 
               
               
                   
               
               
                 AGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGA 
               
               
                   
               
               
                 CGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGT 
               
               
                   
               
               
                 CCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC 
               
               
                   
               
               
                 AGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACC 
               
               
                   
               
               
                 AAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCATCATGCCCAG 
               
               
                   
               
               
                 CACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGA 
               
               
                   
               
               
                 CACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGC 
               
               
                   
               
               
                 CAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATA 
               
               
                   
               
               
                 ATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCA 
               
               
                   
               
               
                 GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCA 
               
               
                   
               
               
                 AGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAA 
               
               
                   
               
               
                 AGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGAT 
               
               
                   
               
               
                 GACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGA 
               
               
                   
               
               
                 CATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC 
               
               
                   
               
               
                 GCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTG 
               
               
                   
               
               
                 GACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGG 
               
               
                   
               
               
                 CTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAATGA 
               
               
                   
               
               
                 &lt;SEQ ID NOS: 21-36&gt; 
                   
               
               
                 SEQ ID NOS: 21-36 show the nucleotide sequences of primers cPD-1 inner F, cPD-1 inner 
                   
               
               
                 R, cPD-L1 inner F, cPD-L1 inner R, cPD-1 5′ GSP, cPD-1 3′ GSP, cPD-L1 5′ GSP, cPD-L1 
               
               
                 3′ GSP, cPD-1-EGFP F, cPD-1-EGFP R, cPD-L1-EGFP F, cPD-L1-EGFP R, cPD-1-Ig, cPD- 
               
               
                 1-Ig R, cPD-L1-Ig F and cPD-L1-Ig R in this order. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 37&gt; 
                   
               
               
                 SEQ ID NO: 37 shows the amino acid sequence (QSLLYSENQKDY) of CDR1 of the VL of 
                   
               
               
                 rat anti-bovine PD-L1 antibody 4G12. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 38&gt; 
                   
               
               
                 SEQ ID NO: 38 shows the amino acid sequence (QSLLYSENQKDY) of CDR3 of the VL of 
                   
               
               
                 rat anti-bovine PD-L1 antibody 4G12, 
               
               
                   
               
               
                 &lt;SEQ ID NO: 39&gt; 
                   
               
               
                 SEQ ID NO: 39 shows the amino acid sequence (GYTFTSNF) of CDR1 of the VH of rat 
                   
               
               
                 anti-bovine PD-L1 antibody 4G12. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 40&gt; 
                   
               
               
                 SEQ ID NO: 40 shows the amino acid sequence (IYPEYGNT) of CDR2 of the VH of rat 
                   
               
               
                 anti-bovine PD-L1 antibody 4G12. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 41&gt; 
                   
               
               
                 SEQ ID NO: 41 shows the amino acid sequence (ASEEAVISLVY) of CDR3 of the VH of rat 
                   
               
               
                 anti-bovine PD-L1 antibody 4G12. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 42&gt; 
                   
               
               
                 SEQ ID NO: 42 shows the amino acid sequence of the CH (CH1—CH3) of ovine antibody 
                   
               
               
                 (IgG1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 43&gt; 
                   
               
               
                 SEQ ID NO: 43 shows the nucleotide sequence of the CH (CH1—CH3) of ovine antibody 
                   
               
               
                 (IgG1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 44&gt; 
                   
               
               
                 SEQ ID NO: 44 he amino acid sequence of the CH (CH1—CH3) of ovine antibody (IgG2). 
                   
               
               
                   
               
               
                 &lt;SEQ ID NO: 45&gt; 
                   
               
               
                 SEQ ID NO: 45 shows the nucleotide sequence of the CH (CH1—-CH3) of ovine antibody 
                   
               
               
                 (IgG2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 46&gt; 
                   
               
               
                 SEQ ID NO: 46 shows the amino acid sequence of the light chain (Ig kappa(CK)) constant 
                   
               
               
                 region of an ovine antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 47&gt; 
                   
               
               
                 SEQ ID NO: 47 shows the nucleotide sequence of the light chain (Ig kappa(CK)) constant 
                   
               
               
                 region of an ovine antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 48&gt; 
                   
               
               
                 SEQ ID NO: 48 shows the amino acid sequence of the light chain (Ig lambda(CL)) constant 
                   
               
               
                 region of an ovine antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 49&gt; 
                   
               
               
                 SEQ ID NO: 49 shows the nucleotide sequence of the light chain (Ig lambda(CL)) constant 
                   
               
               
                 region of an ovine antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 50&gt; 
                   
               
               
                 SEQ ID NO: 50 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG1 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 51&gt; 
                   
               
               
                 SEQ ID NO: 51 shows the nucleotide acid sequence of the CH (CH1—CH3) of porcine 
                   
               
               
                 antibody (IgG1 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 52&gt; 
                   
               
               
                 SEQ ID NO: 52 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG1 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 53&gt; 
                   
               
               
                 SEQ ID NO: 53 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG1 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 54&gt; 
                   
               
               
                 SEQ ID NO: 54 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG2 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 55&gt; 
                   
               
               
                 SEQ ID NO: 55 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG2 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 56&gt; 
                   
               
               
                 SEQ ID NO: 56 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG2 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 57&gt; 
                   
               
               
                 SEQ ID NO: 57 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG2 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 58&gt; 
                   
               
               
                 SEQ ID NO: 58 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 59&gt; 
                   
               
               
                 SEQ ID NO: 59 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 60&gt; 
                   
               
               
                 SEQ ID NO: 60 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG4 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 61&gt; 
                   
               
               
                 SEQ ID NO: 61 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG4 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 62&gt; 
                   
               
               
                 SEQ ID NO: 62 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG4 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 63&gt; 
                   
               
               
                 SEQ ID NO: 63 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG4 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 64&gt; 
                   
               
               
                 SEQ ID NO: 64 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG5 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 65&gt; 
                   
               
               
                 SEQ ID NO: 65 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG5 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 66&gt; 
                   
               
               
                 SEQ ID NO: 66 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG5 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 67&gt; 
                   
               
               
                 SEQ ID NO: 67 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG5 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 68&gt; 
                   
               
               
                 SEQ ID NO: 68 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG6 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 69&gt; 
                   
               
               
                 SEQ ID NO: 69 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG6 a ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 70&gt; 
                   
               
               
                 SEQ ID NO: 70 shows the amino acid sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG6 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 71&gt; 
                   
               
               
                 SEQ ID NO: 71 shows the nucleotide sequence of the CH (CH1—CH3) of porcine antibody 
                   
               
               
                 (IgG6 b ). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 72&gt; 
                   
               
               
                 SEQ ID NO: 72 shows the amino acid sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 73&gt; 
                   
               
               
                 SEQ ID NO: 73 shows the nucleotide sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 74&gt; 
                   
               
               
                 SEQ ID NO: 74 shows the amino acid sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 75&gt; 
                   
               
               
                 SEQ ID NO: 75 shows the nucleotide sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 76&gt; 
                   
               
               
                 SEQ ID NO: 76 shows the amino acid sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 77&gt; 
                   
               
               
                 SEQ ID NO: 77 shows the nucleotide sequence of the CH (CH1—CH3) of a water buffalo 
                   
               
               
                 antibody (estimated to be IgG3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 78&gt; 
                   
               
               
                 SEQ ID NO: 78 shows the amino acid sequence of the light chain (estimated to be Ig lambda) 
                   
               
               
                 constant region (CL) of a water buffalo antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 79&gt; 
                   
               
               
                 SEQ ID NO: 79 shows the nucleotide sequence of the light chain (estimated to be Ig lambda) 
                   
               
               
                 constant region (CL) of a water buffalo antibody. 
               
               
                   
               
               
                 &lt;SEQ ID NO: 80&gt; 
                   
               
               
                 SEQ ID NO: 80 shows the amino acid sequence of the CH (CH1—CH3) of human antibody 
                   
               
               
                 (IgG4 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 81&gt; 
                   
               
               
                 SEQ ID NO: 81 shows the nucleotide sequence of the CH (CH1—CH3) of human antibody 
                   
               
               
                 (IgG4 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 82&gt; 
                   
               
               
                 SEQ ID NO: 82 shows the amino acid sequence of the CH (CH1—CH3) of human antibody 
                   
               
               
                 (IgG4 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 83&gt; 
                   
               
               
                 SEQ ID NO: 83 shows the nucleotide sequence of the CH (CH1-CH3) of human antibody 
                   
               
               
                 (IgG4 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 84&gt; 
                   
               
               
                 SEQ ID NO: 84 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 85&gt; 
                   
               
               
                 SEQ ID NO: 85 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 86&gt; 
                   
               
               
                 SEQ ID NO: 86 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 87&gt; 
                   
               
               
                 SEQ ID NO: 87 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 88&gt; 
                   
               
               
                 SEQ ID NO: 88 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 89&gt; 
                   
               
               
                 SEQ ID NO: 89 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 90&gt; 
                   
               
               
                 SEQ ID NO: 90 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG3 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 91&gt; 
                   
               
               
                 SEQ ID NO: 91 shows the amino acid sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG3 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 92&gt; 
                   
               
               
                 SEQ ID NO: 92 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 93&gt; 
                   
               
               
                 SEQ ID NO: 93 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 94&gt; 
                   
               
               
                 SEQ ID NO: 94 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG1 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 95&gt; 
                   
               
               
                 SEQ ID NO: 95 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 96&gt; 
                   
               
               
                 SEQ ID NO: 96 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 97&gt; 
                   
               
               
                 SEQ ID NO: 97 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG2 variant 3). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 98&gt; 
                   
               
               
                 SEQ ID NO: 98 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG3 variant 1). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 99&gt; 
                   
               
               
                 SEQ ID NO: 99 shows the nucleotide sequence of the CH (CH1—CH3) of bovine antibody 
                   
               
               
                 (IgG3 variant 2). 
               
               
                   
               
               
                 &lt;SEQ ID NO: 100&gt; 
                   
               
               
                 SEQ ID NO: 100 shows the amino acid sequence of the CL of a bovine antibody (bovine Ig 
                   
               
               
                 lambda, GenBank: X62917). 
               
               
                 QPKSPPSVTLFPPSTEELNGNKATLVCLISDFYPGSVTVVWKADGSTITRNVETTRASK 
               
               
                   
               
               
                 QSNSKYAASSYLSLTSSDWKSKGSYSCEVTHEGSTVTKTVKPSECS 
               
               
                   
               
               
                 &lt;SEQ ID NO: 101&gt; 
                   
               
               
                 SEQ ID NO: 101 shows the nucleotide sequence of the CL of a bovine antibody (bovine Ig 
                   
               
               
                 lambda, GenBank: X62917). 
               
               
                 CAGCCCAAGTCCCCACCCTCGGTCACCCTGTTCCCGCCCTCCACGGAGGAGCTCA 
               
               
                   
               
               
                 ACGGCAACAAGGCCACCCTGGTGTGTCTCATCAGCGACTTCTACCCGGGTAGCGT 
               
               
                   
               
               
                 GACCGTGGTCTGGAAGGCAGACGGCAGCACCATCACCCGCAACGTGGAGACCAC 
               
               
                   
               
               
                 CCGGGCCTCCAAACAGAGCAACAGCAAGTACGCGGCCAGCAGCTACCTGAGCCT 
               
               
                   
               
               
                 GACGAGCAGCGACTGGAAATCGAAAGGCAGTTACAGCTGCGAGGTCACGCACGA 
               
               
                   
               
               
                 GGGGAGCACCGTGACGAAGACAGTGAAGCCCTCAGAGTGTTCTTAG 
               
               
                   
               
               
                 The nucleotide sequence of SEQ ID NO: 101 after codon optimization is shown in &lt;SEQ ID 
               
               
                 NO: 104&gt;. 
               
               
                 CAGCCTAAGAGTCCTCCTTCTGTAACACTCTTTCCCCCCTCTACCGAGGAACTCAA 
               
               
                   
               
               
                 CGGCAATAAAGCTACCTTGGTTTGCCTTATTTCTGATTTCTACCCCGGGTCTGTGAC 
               
               
                   
               
               
                 CGTGGTGTGGAAAGCTGATGGGTCCACCATTACTCGGAATGTGGAAACCACCCGG 
               
               
                   
               
               
                 GCTTCTAAGCAGTCCAACTCTAAATACGCAGCATCCTCCTATTTGAGTCTTACTAGT 
               
               
                   
               
               
                 AGTGACTGGAAGTCAAAGGGTAGTTACAGTTGCGAAGTCACACATGAAGGTTCA 
               
               
                   
               
               
                 ACAGTGACAAAGACAGTCAAGCCCTCAGAGTGCTCATAG 
               
               
                   
               
               
                 &lt;SEQ ID NO: 102&gt; 
                   
               
               
                 SEQ ID NO: 102 shows the amino acid sequence of the CH of a bovine antibody (bovine 
                   
               
               
                 IgG1, modified from GenBank: X62916). The sites of mutation are underlined. Amino acid 
               
               
                 numbers and mutations: 113E→P, 114L→V, 115P→A, 116G→deletion, 209A→S, 210P→S 
               
               
                 ASTTAPKVYPLSSCCGDKSSSTVTLGCLVSSYMPEPVTVTWNSGALKSGVHTFPAVL 
               
               
                   
               
               
                 QSSGLYSLSSMVTVPGSTSGQTFTCNVAHPASSTKVDKAVDPTCKPSPCDCCPPP PVA   
               
               
                   
               
               
                 GPSVFIFPPKPKDTLTISGTPEVTCVVVDVGHDDPEVKFSWFVDDVEVNTATTKPREE 
               
               
                   
               
               
                 QFNSTYRVVSALRIQHQDWTGGKEFKCKVHNEGLP SS IVRTISRTKGPAREPQVYVLA 
               
               
                   
               
               
                 PPQEELSKSTVSLTCMVTSFYPDYIAVEWQRNGQPESEDKYGTTPPQLDADSSYFLYS 
               
               
                   
               
               
                 KLRVDRNSWQEGDTYTCVVMHEALHNHYTQKSTSKSAGK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 103&gt; 
                   
               
               
                 SEQ ID NO: 103 shows the nucleotide sequence (after codon optimization) of the CH of a 
                   
               
               
                 bovine antibody (bovine IgG1, modified from GenBank: X62916). 
               
               
                 GCTAGCACAACTGCTCCTAAGGTGTACCCCCTGAGCTCTTGCTGCGGCGACAAGT 
               
               
                   
               
               
                 CTAGCAGCACCGTGACCCTCGGATGCCTCGTCAGCAGCTATATGCCTGAGCCAGTT 
               
               
                   
               
               
                 ACAGTGACATGGAATTCTGGTGCCCTTAAGTCCGGCGTCCATACCTTCCCTGCTGT 
               
               
                   
               
               
                 GCTGCAGTCCTCTGGCCTGTACAGTTTGTCCTCTATGGTGACAGTACCCGGTTCCA 
               
               
                   
               
               
                 CCTCCGGACAGACCTTTACCTGTAATGTGGCTCATCCCGCCTCCTCCACAAAGGTG 
               
               
                   
               
               
                 GATAAGGCTGTTGACCCTACCTGTAAACCCAGTCCATGCGACTGCTGTCCCCCCCC 
               
               
                   
               
               
                 TCCAGTTGCCGGACCCTCAGTCTTTATTTTCCCACCCAAACCCAAAGACACCCTGA 
               
               
                   
               
               
                 CAATCTCTGGAACACCAGAAGTCACCTGCGTCGTCGTGGATGTGGGCCACGACGA 
               
               
                   
               
               
                 TCCTGAGGTAAAATTCTCATGGTTCGTCGACGATGTGGAAGTGAATACAGCTACTA 
               
               
                   
               
               
                 CAAAACCTCGCGAAGAGCAGTTTAACTCTACCTATCGAGTGGTTTCTGCTTTGCGG 
               
               
                   
               
               
                 ATTCAGCATCAGGATTGGACAGGCGGCAAAGAGTTTAAATGTAAAGTCCATAACG 
               
               
                   
               
               
                 AGGGACTTCCTTCTAGTATCGTGCGCACTATCAGTAGAACTAAAGGGCCTGCTCGG 
               
               
                   
               
               
                 GAACCTCAGGTGTACGTCCTGGCACCTCCACAGGAAGAGCTGAGTAAGTCTACAG 
               
               
                   
               
               
                 TTTCTCTGACTTGTATGGTAACATCTTTTTATCCAGATTACATCGCAGTTGAATGGC 
               
               
                   
               
               
                 AGAGGAACGGGCAGCCAGAGAGTGAGGATAAGTACGGGACTACTCCACCACAGC 
               
               
                   
               
               
                 TGGACGCAGACTCAAGTTACTTCCTGTACTCAAAGCTGAGGGTTGACAGAAACTC 
               
               
                   
               
               
                 ATGGCAGGAGGGGGACACTTACACTTGCGTAGTTATGCACGAGGCACTTCACAAC 
               
               
                   
               
               
                 CACTACACTCAGAAGAGTACTTCAAAGAGTGCAGGGAAGTAA 
               
               
                   
               
               
                 &lt;SEQ ID NO: 105&gt; 
                   
               
               
                 SEQ ID NO: 105 shows the amino acid sequence of a chimeric light chain consisting of the 
                   
               
               
                 VL of rat anti-bovine PD-L1 antibody and the CL of a bovine antibody. 
               
               
                 MESQTHVLISLLLSVSGTYGDIAITQSPSSVAVSVGETVTLSCKSSQSLLYSENQKDYL 
               
               
                   
               
               
                 GWYQQKPGQTPKPLIYWATNRHTGVPDRFTGSGSGTDFTLIISSVQAEDLADYYCGQ 
               
               
                   
               
               
                 YLVYPFTFGPGTKLELKQPKSPPSVTLFPPSTEELNGNKATLVCLISDFYPGSVTVVWK 
               
               
                   
               
               
                 ADGSTITRNVETTRASKQSNSKYAASSYLSLTSSDWKSKGSYSCEVTHEGSTVTKTV 
               
               
                   
               
               
                 KPSECS 
               
               
                   
               
               
                 &lt;SEQ ID NO: 106&gt; 
                   
               
               
                 SEQ ID NO: 106 shows the amino acid sequence of a chimeric heavy chain consisting of the 
                   
               
               
                 VH of rat anti-bovine PD-L1 antibody and the CH of a bovine antibody (bovine IgG1, 
               
               
                 modified from GenBank: X62916). 
               
               
                 MGWSQIILFLVAAATCVHSQVQLQQSGAELVKPGSSVKISCKASGYTFTSNFMHWVK 
               
               
                   
               
               
                 QQPGNGLEWIGWIYPEYGNTKYNQKFDGKATLTADKSSSTAYMQLSSLTSEDSAVYF 
               
               
                   
               
               
                 CASEEAVISLYWGQGTLVTVSSASTTAPKVYPLSSCCGDKSSSTVTLGCLVSSYMPE 
               
               
                   
               
               
                 PVTVTWNSGALKSGVHTFPAVLQSSGLYSLSSMVTVPGSTSGQTFTCNVAHPASSTKV 
               
               
                   
               
               
                 DKAVDPTCKPSPCDCCPPPPVAGPSVFIFFPKPKDTLTISGTPEVTCVVVDVGHDDPEV 
               
               
                   
               
               
                 KFSWFVDDVEVNTATTKPREEQFNSTYRVVSALRIQHQDWTGGKEFKCKVHNEGLP 
               
               
                   
               
               
                 SSIVRTISRTKGPAREPQVYVLAPPQEELSKSTVSLTCMVTSFYPDYIAVEWQRNGQPE 
               
               
                   
               
               
                 SEDKYGTTPPQLDADSSYFLYSKLRVDRNSWQEGDTYTCVVMHEALHNHYTQKSTS 
               
               
                   
               
               
                 KSAGK 
               
               
                   
               
               
                 &lt;SEQ ID NO: 107&gt; 
                   
               
               
                 This sequence shows the nucleotide sequence (after codon optimization) of a chimeric light 
                   
               
               
                 chain consisting of the VL of rat anti-bovine PD-L1 antibody and the CL of a bovine 
               
               
                 antibody. 
               
               
                 ATGGAATCTCAAACTCATGTTTTGATTTCATTACTTCTGAGTGTTTCCGGAACCTAC 
               
               
                   
               
               
                 GGTGATATCGCTATCACTCAATCTCCCTCCTCTGTTGCTGTGTCTGTGGGCGAAAC 
               
               
                   
               
               
                 CGTTACCCTGTCCTGCAAGTCCAGTCAGTCTCTTCTCTACTCCGAGAATCAAAAGG 
               
               
                   
               
               
                 ACTACCTGGGCTGGTACCAACAGAAGCCCGGCCAGACCCCAAAGCCACTGATATA 
               
               
                   
               
               
                 CTGGGCAACCAACCAACGGCACACCGGAGTGCCCGACAGGTTCACAGGCAGTGGATC 
               
               
                   
               
               
                 TGGCACCGACTTTACCTTGATCATTTCAAGCGTGCAGGCTGAAGATCTGGCCGACT 
               
               
                   
               
               
                 ACTACTGTGGTCAGTATCTGGTGTATCCTTTCACTTTCGGGCCAGGGACAAAACTC 
               
               
                   
               
               
                 GAGCTCAAACAGCCTAAGAGTCCTCCTTCTGTAACACTCTTTCCCCCCTCTACCGA 
               
               
                   
               
               
                 GGAACTCAACGGCAATAAAGCTACCTTGGTTTGCCTTATTTCTGATTTCTACCCCG 
               
               
                   
               
               
                 GGTCTGTGACCGTGGTGTGGAAAGCTGATGGGTCCACCATTACTCGGAATGTGGA 
               
               
                   
               
               
                 AACCACCCGGGCTTCTAAGCAGTCCAACTCTAAATACGCAGCATCCTCCTATTTGA 
               
               
                   
               
               
                 GTCTTACTAGTAGTGACTGGAAGTCAAAGGGTAGTTACAGTTGCGAAGTCACACA 
               
               
                   
               
               
                 TGAAGGTTCAACAGTGACAAAGACAGTCAAGCCCTCAGAGTGCTCATAG 
               
               
                   
               
               
                 &lt;SEQ ID NO: 108&gt; 
                   
               
               
                 This sequence shows the nucleotide sequence (after codon optimization) of a chimeric heavy 
                   
               
               
                 chain consisting of the VH of rat anti-bovine PD-L1 antibody and the CH of a bovine 
               
               
                 antibody (bovine IgG1, modified from GenBank: X62916). 
               
               
                 ATGGGGTGGTCCCAGATTATATTGTTCCTCGTCGCCGCCGCCACTTGCGTACACAG 
               
               
                   
               
               
                 CCAAGTGCAACTTCAACAAAGCGGTGCAGAACTGGTAAAGCCCGGTAGCTCTGT 
               
               
                   
               
               
                 GAAAATATCCTGTAAAGCCAGTGGCTACACATTTACCAGCAACTTTAGCACTGGG 
               
               
                   
               
               
                 TGAAGCAACAGCCCGGAAATGGCTTGGAGTGGATTGGCTGGATCTATCCCGAATAT 
               
               
                   
               
               
                 GGTAACACCAAGTATAATCAGAAGTTCGACGGTAAGGCCACCCTCACCGCCGATA 
               
               
                   
               
               
                 AGTCATCCTCCACCGCCTATATGCAGCTCAGCAGCCTGACCAGCGAGGATTCCGCT 
               
               
                   
               
               
                 GTGTACTTCTGTGCCAGCGAAGAGGCTGTGTGATCTCATTGGTGTATTGGGGACAGG 
               
               
                   
               
               
                 GCACCCTCGTCACCGTGTCCAGCGCTAGCACAACTGCTCCTAAGGTGTACCCCCT 
               
               
                   
               
               
                 GAGCTCTTGCTGCGGCGACAAGTCTAGCAGCACCGTGACCCTCGGATGCCTCGTC 
               
               
                   
               
               
                 AGCAGCTATAGCCTGAGCCAGTTACAGTGACATGGAATTCTGGTGCCCTTAAGTC 
               
               
                   
               
               
                 CGGCGTCCATACCTTCCCTGCTGTGCTGCAGTCCTCTGGCCTGTACAGTTTGTCCT 
               
               
                   
               
               
                 CTATGGTGACAGTACCCGGTTCCACCTCCGGACAGACCTTTACCTGTAATGTGGCT 
               
               
                   
               
               
                 CATCCCGCCTCCTCCACAAAGGTGGATAAGGCTGTTGACCCTACCTGTAAACCCA 
               
               
                   
               
               
                 GTCCATGCGACTGCTGTCCCCCCCCTCCAGTTGCCGGACCCTCAGTCTTTATTTTC 
               
               
                   
               
               
                 CCACCCAAACCCAAAGACACCCTGACAATCTCTGGAACACCAGAAGTCACCTGC 
               
               
                   
               
               
                 GTCGTCGTGGATGTGGGCCACGACGATCCTGAGGTAAAATTCTCATGGTTCGTCGA 
               
               
                   
               
               
                 CGATGTGGAAGTGAATACAGCTACTACAAAACCTCGCGAAGAGCAGTTTAACTCT 
               
               
                   
               
               
                 ACCTATCGAGTGGTTTCTGCTTTGCGGATTCAGCATCAGGATTGGACAGGCGGCAA 
               
               
                   
               
               
                 AGAGTTTAAATGTAAAGTCCATAACGAGGGACTTCCTTCTAGTATCGTGCGCACTA 
               
               
                   
               
               
                 TCAGTAGAACTAAAGGGCCTGCTCGGGAACCTCAGGTGTACGTCCTGGCACCTCC 
               
               
                   
               
               
                 ACAGGAAGAGCTGAGTAAGTCTACAGTTTCTCTGACTTGTATGGTAACATCTTTTT 
               
               
                   
               
               
                 ATCCAGATTACATCGCAGTTGAATGGCAGAGGAACGGGCAGCCAGAGAGTGAGG 
               
               
                   
               
               
                 ATAAGTACGGGACTACTCCACCACAGCTGGACGCAGACTCAAGTTACTTCCTGTA 
               
               
                   
               
               
                 CTCAAAGCTGAGGGTTGACAGAAACTCATGGCAGGAGGGGGACACTTACACTTG 
               
               
                   
               
               
                 CGTAGTTATGCACGAGGCACTTCACAACCACTACACTCAGAAGAGTACTTCAAAG 
               
               
                   
               
               
                 AGTGCAGGGAAGTAA 
               
               
                   
               
               
                 &lt;SEQ ID NOS: 109-132&gt; 
                   
               
               
                 SEQ ID NOS: 109-132 show the nucleotide sequences of primers ovPD-L1 CDS F, ovPD-L1 
                   
               
               
                 CDS R, poPD-L1 CDS F, poPD-L1 CDS R, buPD-L1 CDS F1, buPD-L1 CDS R1, buPD-L1 
               
               
                 CDS F2, buPD-L1 CDS R2, ovPD-1-EGFP F, ovPD-1-EGFP R, ovPD-L1-EGFP F, ovPD- 
               
               
                 L1-EGFP R, poPD-1-EGFP F, poPD-1-EGFP R, poPD-L1-EGFP F, poPD-L1-EGFP R, 
               
               
                 ovPD-L1-Ig F, ovPD-L1-Ig R, poPD-L1-Ig F, poPD-L1-Ig R, cCD80-Ig F, cCD80-Ig R, cPD- 
               
               
                 L1-His F and cPD-L1-His R in this order. 
               
               
                   
               
               
                 &lt;SEQ ID NOS: 133-138&gt; 
                   
               
               
                 SEQ ID NOS: 133-138 show nucleotide sequences of primers boPD-1-myc F, boPD-1-myc 
                   
               
               
                 R, boPD-L1-EGFP F, boPD-L1-EGFP R, boPD-L1-Ig F and boPD-L1-Ig R in this order.