A METHOD FOR IN VITRO MOLECULAR EVOLUTION OF ANTIBODY FUNCTION

摘要

The present invention provides a method for producing a polynucleotide sequence encoding an antibody variable domain, the variable domain comprising complementarity-determining regions (CDRs) located within a selected framework (the 'master framework'), the method comprising the steps of (a) providing at least one nucleic acid molecule encoding one or more CDRs and associated framework regions (the 'original framework'), (b) amplifying at least one CDR-encoding portion of the nucleic acid molecule(s) of step (a) using one or more pairs of oligonucleotides as amplification primers and (c) assembling a polynucleotide sequence encoding an antibody variable domain by combining the amplified CDR-encoding nucleotide sequences produced in step (b) with nucleotide sequences encoding said master framework, wherein the oligonucleotide primers of step (b) comprise nucleotide sequences which differ from the corresponding nucleotide sequences encoding said master framework. The invention further provides an antibody library, such as a phage display library, and methods of making the same.