Anti-ephb2 antibodies and methods using same

Abstract

The invention provides anti-EphB2 antibodies, immunoconjugates, and compositions comprising and methods of using these antibodies and immunoconjugates.

1 . An isolated polynucleotide encoding an anti-EphB2 antibody comprising: at least one, two, three, four, five, and/or six hypervariable region (HVR) sequences selected from the group consisting of:
 (a) HVR-L1 comprising sequence KSSQSLLNSGNQENYLA (SEQ ID NO:1);   (b) HVR-L2 comprising sequence GASTRES (SEQ ID NO:2);   (c) HVR-L3 comprising sequence QNDHSYPFT (SEQ ID NO:3);   (d) HVR-H1 comprising sequence SYWMH (SEQ ID NO:4);   (e) HVR-H2 comprising sequence FINPSTGYTDYNQKFKD (SEQ ID NO:5); and   (f) HVR-H3 comprising sequence RLKLLRYAMDY (SEQ ID NO:6),   wherein the antibody specifically binds human EphB2.   
     
     
         2 . The polynucleotide of  claim 1 , wherein the antibody comprises a light chain comprising at least one, at least two, or all three of HVR sequences selected from the group consisting of KSSQSLLNSGNQENYLA (SEQ ID NO:1), GASTRES (SEQ ID NO:2), and QNDHSYPFT (SEQ ID NO:3). 
     
     
         3 . The polynucleotide of  claim 1  or  2 , wherein the antibody comprises a heavy chain comprising at least one, at least two or all three of HVR sequences selected from the group consisting of SYWMH (SEQ ID NO:4), FINPSTGYTDYNQKFKD (SEQ ID NO:5), and RLKLLRYAMDY (SEQ ID NO:6). 
     
     
         4 . The polynucleotide of  claim 1 , wherein the antibody comprises
 (a) a heavy chain comprising at least one, at least two or all three of HVR sequences selected from the group consisting of SYWMH (SEQ ID NO:4), FINPSTGYTDYNQKFKD (SEQ ID NO:5), and RLKLLRYAMDY (SEQ ID NO:6); and   (b) a light chain comprising at least one, at least two or all three of CDR sequences selected from the group consisting of KSSQSLLNSGNQENYLA (SEQ ID NO:1), GASTRES (SEQ ID NO:2), and QNDHSYPFT (SEQ ID NO:3).   
     
     
         5 . The polynucleotide of  claim 1 , wherein the antibody comprises a light chain variable domain having the sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 7) 
                 
                   DIVMTQSPSSLSVSAGEKVTMNCKSSQSLLNSGNQENYLAWYQQKPG 
                 
                     
                 
                   QPPKLLIYGASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYC 
                 
                     
                 
                   QNDHSYPFTFGAGTKVEIKR. 
                 
             
                
                
                
                
                
                
               
            
           
         
       
     
     
         6 . The polynucleotide of  claim 1  or  5 , wherein the antibody comprises a heavy chain variable domain having the sequence: 
       
         
           
                 
               
                   (SEQ ID NO: 8 
                 
                   QVQLQQSGAELAKPGASVKMSCKASGYTFTSYWMHWVKQRPGQGLEW 
                 
                     
                 
                   IGFINPSTGYTDYNQKFKDKATLTVKSSNTAYMQLSRLTSEDSAVYY 
                 
                     
                 
                   CTRRLKLLRYAMDYWGQGTTLTVSA. 
                 
             
                
                
                
                
                
                
               
            
           
         
       
     
     
         7 . An isolated polynucleotide encoding an antibody comprising at least one hypervariable sequence comprising a sequence selected from the group consisting of HVR-L1, HVR-L2, HVR-L3, HVR-H1, HVR-H2, and HVR-H3 of the antibody produced by hybridoma cell line having American Tissue Type Culture (ATCC) No. PTA-6606, wherein said isolated antibody specifically binds human EphB2. 
     
     
         8 . An isolated polynucleotide encoding an antibody comprising heavy and/or light chain variable domain of the antibody produced by hybridoma cell line having American Tissue Type Culture (ATCC) No. PTA-6606, wherein said isolated antibody specifically binds human EphB2. 
     
     
         9 . An isolated polynucleotide encoding an antibody that binds to a same epitope on human EphB2 as the antibody produced by hybridoma cell line having American Tissue Type Culture (ATCC) No. PTA-6606. 
     
     
         10 . An isolated polynucleotide encoding an antibody that competes with an antibody produced by hybridoma cell line having American Tissue Type Culture (ATCC) No. PTA-6606 for binding to human EphB2. 
     
     
         11 . An isolated polynucleotide encoding an antibody encoded by antibody coding sequences of hybridoma cell line having American Tissue Type Culture (ATCC) No. PTA-6606 
     
     
         12 . The polynucleotide of  claim 1 ,  9  or  10 , wherein the antibody is a monoclonal antibody. 
     
     
         13 . The polynucleotide of  claim 1 ,  9  or  10 , wherein the antibody is selected from the group consisting of a chimeric antibody, a humanized antibody, an affinity matured antibody, a human antibody, and a bispecific antibody. 
     
     
         14 . The polynucleotide of  claim 1 ,  9  or  10 , wherein the antibody is an antibody fragment. 
     
     
         15 . A vector comprising the polynucleotide of  claim 1 ,  9  or  10 . 
     
     
         16 . The vector of  claim 15 , wherein the vector is an expression vector. 
     
     
         17 . A host cell comprising a vector of  claim 15  or  16 . 
     
     
         18 . The host cell of  claim 17 , wherein the host cell is prokaryotic. 
     
     
         19 . The host cell of  claim 17 , wherein the host cell is eukaryotic. 
     
     
         20 . The host cell of  claim 17 , wherein the host cell is mammalian. 
     
     
         21 . A method for making an anti-EphB2 antibody, said method comprising culturing a host cell comprising polynucleotide encoding the antibody of  claim 1 ,  9  or  10  so that the polynucleotide is expressed, and optionally, recovering the antibody from the culture. 
     
     
         22 . A method for making an anti-EphB2 immunoconjugate, said method comprising culturing a host cell comprising polynucleotide encoding the antibody of  claim 1 ,  9  or  10  so that the polynucleotide is expressed, and optionally, recovering the antibody from the culture. 
     
     
         23 . The method of  claim 21  or  22 , wherein the host cell is prokaryotic. 
     
     
         24 . The method of  claim 21  or  22 , wherein the host cell is eukaryotic. 
     
     
         25 . The method of  claim 21 , further comprising recovering the antibody from the host cell culture. 
     
     
         26 . The method of  claim 25 , where in the antibody is recovered from the host cell culture medium. 
     
     
         27 . The method of  claim 25 , further comprising combining the recovered antibody with a pharmaceutically acceptable carrier, excipient, or carrier to prepare a pharmaceutical formulation comprising the antibody. 
     
     
         28 . A composition comprising a polynucleotide of  claim 14 ,  15  or  16 . 
     
     
         29 . The composition of  claim 28 , wherein the composition further comprises a carrier.