Methods of generating libraries and uses thereof

摘要

This invention relates to methods for the generation of humanized antibodies, particularly a humanized antibody heavy chain protein and a humanized antibody light chain protein. The method comprises using cells that express or can be induced to express Activation Induced Cytidine Deaminase (AID).

主权项

1. A method of preparing a library of antibody heavy chain-encoding polynucleotides, which method comprises:
(a) preparing a first library of nucleic acid sequences, wherein each nucleic acid sequence encodes a different CDR3 domain of an antibody heavy chain variable region, (b) preparing a second library of nucleic acid sequences, wherein each nucleic acid sequence encodes a template antibody heavy chain variable region domain that lacks an endogenous CDR3 domain, (c) preparing a third library of nucleic acid sequences, wherein each nucleic acid sequence encodes an antibody heavy chain constant region template, (d) operably linking the nucleic acid sequences of the libraries of (a), (b), and (c) in an expression vector to produce a fourth library of antibody heavy chain-encoding polynucleotides, (e) transforming host cells with the library of antibody heavy chain-encoding polynucleotides produced in step (d), and (f) expressing Activation Induced Cytidine Deaminase (AID) in the host cells, wherein AID induces one or more mutations in the library of antibody heavy chain-encoding polynucleotides transformed into the host cells in step (e) to provide a fifth library of heavy chain-encoding polynucleotides.