Patent ID: 9090994
Filing Date: 2015-07-28
Classification: C07K,C40B

Abstract:
1. A method of producing a humanized antibody or an antigen binding fragment thereof for a non-human antibody to an antigen, comprising: (1) constructing a heavy chain variable domain (VH) framework (FR) library comprising nucleic acids encoding a diversity of VHs, each of the VHs comprising FR1, FR2, FR3 and FR4 independently selected from corresponding FRs of a first number of homologous human germline VHs, and corresponding complementarity determining regions (CDRs) of the VH of the non-human antibody; (2) constructing a light chain variable domain (VL) FR library comprising nucleic acids encoding a diversity of VLs, each of the VLs comprising FR1, FR2, FR3 and FR4 independently selected from corresponding FRs of a second number of homologous human germline VLs, and corresponding CDRs of the VL of the non-human antibody; (3) constructing a framework-assembly library comprising nucleic acids encoding a diversity of Fabs or fragments thereof, each of the Fabs or the fragments thereof comprising a VH encoded by a nucleic acid in the VH FR library and a VL encoded by a nucleic acid in the VL FR library; (4) expressing the framework-assembly library in host cells; (5) identifying from the expressed framework-assembly library an Fab or a fragment thereof that binds to the antigen; (6) identifying the VH and VL in the identified Fab or the fragment thereof as a humanized VH and a humanized VL, respectively, for the humanized antibody or the antigen binding fragment thereof; and (7) producing the humanized antibody or the antigen binding fragment thereof comprising the identified humanized VH and the humanized VL, wherein the sequences of FR1, FR2, FR3, and FR4 in the VH FR library and VL FR library are natural human germline sequences.