Construction of a fully synthetic human phage display antibody library

Objective:To construct a fully synthetic human phage display antibody library.Methods:The frameworks of human antibody germline genes and semi-random CDR3s(complementarity determining region 3) were synthesized artificially.The antibody genes were synthesized by SOE-PCR(splicing overlap extension PCR).Then the antibody genes and the phage display vector were digested by restriction endonucleases SfiⅠ and NotⅠ.The recombinant phagemid was electroporated into E.coli TG1.Results and conclusion:As long as the diversity of antibody library was ensured,the antibody genes could be obtained expediently by the improved method.A fully synthetic human phage display antibody library of 2×10~(9)was constructed through numerous electro-transformations.The quality of the library was proven to be fine by colony PCR and sequencing.It established a base for screening human therapeutic antibody.