Patent ID: 8748356
Filing Date: 2014-06-10
Classification: C07K

Abstract:
1. A method of making human-adapted antibodies comprising selecting a human framework based on overall similarity to a non-human antibody variable region comprising the steps of: a) obtaining a polypeptide sequence of a non-human antibody variable region; b) delineating complementarity determining regions 1, 2, and 3 (CDR1, CDR2, and CDR3) and framework regions 1, 2, 3, and 4 (FR1, FR2, FR3 and FR4) in the polypeptide sequence of the non-human antibody variable region; c) providing a library of polypeptide sequences encoded by human variable region (V) and joining region (J) germline genes; d) selecting a first subset of polypeptide sequences encoded by the human V germline genes from the library of polypeptide sequences provided in step b that have maximum similarities to over the entire length of the polypeptide sequence of the non-human antibody variable region; e) selecting from the first subset of polypeptide sequences those polypeptide sequences that are most similar at CDR1 and CDR2 based on CDR1 and CDR2 amino acid sequence and length when compared to the non-human antibody variable region when the polypeptide sequence selected in step c is a polypeptide sequence encoding a heavy chain variable region, and those polypeptide sequences that are most similar at CDR1, CDR2, and CDR3 based on CDR1, CDR2, and CDR3 amino acid sequence and length when the polypeptide sequence identified in step c is a polypeptide sequence encoding a light chain variable region; f) selecting a second subset of polypeptide sequences encoded by the human J germline genes from the library of polypeptide sequences provided in step b that have maximum similarities to the polypeptide sequence encoded by a framework region 4 (FR4) of the non-human antibody variable region; g) generating polypeptide sequences of human antibody variable regions that show maximum homology to the non-human antibody variable region by joining the first subset of polypeptide sequences obtained in step e with the second subset of polypeptide sequences obtained in step f; h) transferring CDRs of the non-human antibody variable region into the polypeptide sequences of human antibody variable regions generated in step g to form human-adapted antibody variable regions; and i) expressing the human-adapted antibody variable regions to make the human-adapted antibodies.