Specific binding proteins and uses thereof

Abstract

The present invention relates to specific binding members, particularly antibodies and fragments thereof, which bind to amplified epidermal growth factor receptor (EGFR) and to the de2-7 EGFR truncation of the EGFR. In particular, the epitope recognized by the specific binding members, particularly antibodies and fragments thereof, is enhanced or evident upon aberrant post-translational modification. These specific binding members are useful in the diagnosis and treatment of cancer. The binding members of the present invention may also be used in therapy in combination with chemotherapeutics or anti-cancer agents and/or with other antibodies or fragments thereof.

What is claimed is: 
     
         1 . A method of treating a human subject having a tumor, the method comprising administering to the subject a therapeutically effective amount of an isolated anti-Epidermal Growth Factor Receptor (EGFR) antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 164, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 166. 
     
     
         2 . The method according to  claim 1 , wherein the heavy chain comprises the constant region set forth in SEQ ID NO: 43. 
     
     
         3 . The method according to  claim 1 , wherein the light chain comprises the constant region set forth in SEQ ID NO: 48. 
     
     
         4 . The method according to  claim 3 , wherein the antibody is an IgG isotype. 
     
     
         5 . The method according to  claim 4 , wherein the IgG isotype is an IgG1 isotype. 
     
     
         6 . The method according to  claim 1 , wherein the tumor is a solid tumor. 
     
     
         7 . The method according to  claim 6 , wherein the solid tumor is selected from the group consisting of a breast tumor, a lung tumor, a prostate tumor, a bladder tumor, a head tumor, and a neck tumor. 
     
     
         8 . The method according to  claim 6 , wherein the solid tumor is a glioma. 
     
     
         9 . The method according to  claim 6 , wherein the solid tumor is an epithelial tumor. 
     
     
         10 . The method according to  claim 1 , wherein the antibody is conjugated to a cytotoxic agent. 
     
     
         11 . The method according to  claim 1 , wherein the antibody is administered in combination with temozolomide. 
     
     
         12 . A method of treating a human subject having cancer, comprising administering to the subject a therapeutically effective amount of an immunoconjugate comprising an anti-EGFR antibody conjugated to a cytotoxic agent, wherein the anti-EGFR antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 164, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 166. 
     
     
         13 . The method according to  claim 12 , wherein the heavy chain comprises the constant region set forth in SEQ ID NO: 43. 
     
     
         14 . The method according to  claim 12 , wherein the light chain comprises the constant region set forth in SEQ ID NO: 48. 
     
     
         15 . The method according to  claim 14 , wherein the antibody is an IgG isotype. 
     
     
         16 . The method according to  claim 15 , wherein the IgG isotype is an IgG1 isotype. 
     
     
         17 . The method according to  claim 12 , wherein the immunoconjugate is administered in combination with an anti-cancer agent. 
     
     
         18 . The method according to  claim 17 , wherein the anti-cancer agent is temozolomide. 
     
     
         19 . A method of treating a human subject having a tumor with amplified EGFR, the method comprising administering to the subject an immunoconjugate comprising an anti-EGFR antibody conjugated to a cytotoxic agent, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 164, comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 166, and is an IgG1 isotype. 
     
     
         20 . The method according to  claim 19 , wherein the tumor is selected from the group consisting of a medulloblastoma, a meningioma, a neoplastic astrocytoma, and a neoplastic arteriovenous malformation. 
     
     
         21 . The method according to  claim 19 , wherein the tumor is a glioblastoma. 
     
     
         22 . The method according to  claim 21 , wherein the immunoconjugate is administered in combination with an anti-cancer agent. 
     
     
         23 . The method according to  claim 21 , wherein the anti-cancer agent is temozolomide. 
     
     
         24 . The method according to  claim 19 , wherein the heavy chain comprises the constant region set forth in SEQ ID NO: 43. 
     
     
         25 . The method according to  claim 19 , wherein the light chain comprises the constant region set forth in SEQ ID NO: 48. 
     
     
         26 . The method according to  claim 25 , wherein the tumor is an epithelial tumor. 
     
     
         27 . The method according to  claim 25 , wherein the tumor is a glioblastoma. 
     
     
         28 . The method according to  claim 27 , wherein the immunoconjugate is administered in combination with an anti-cancer agent. 
     
     
         29 . The method according to  claim 28 , wherein the anti-cancer agent is temozolomide. 
     
     
         30 . An isolated antibody capable of binding EGFR on tumors containing amplifications of the EGFR gene, wherein cells of said tumors contain multiple copies of the EGFR gene, and on tumors that express the truncated version of the EGFR receptor de2-7, wherein said antibody does not bind to the de2-7 EGFR junctional peptide consisting of the amino acid sequence of SEQ ID NO:13, wherein said antibody binds to an epitope within the sequence of residues 287-302 (SEQ ID NO:14) of human wild-type EGFR, and wherein said antibody does not comprise a heavy chain variable region sequence having the amino acid sequence set forth in SEQ ID NO:2 and does not comprise a light chain variable region sequence having the amino acid sequence set forth in SEQ ID NO:4.