Construction and screening of phage antibody library for scFv fragment from recovery phase patient infected with H5N1 virus

Objective To construct and screen the phage antibody library for seFv fragment from recovery phase patient infected with H5N1 virus.Methods Total RNA was extractod from peripheral blood lymphocytes from a patient infected with H5N1 virus and was transformed to cDNA by reverse transcription.The variable domains of heavy and light chains were amplified by PCR with a set of specific primer of human IgG.The scFv gene fragments linked VH and VL with synthesized scFv linker were cloned into phagemid pCANTAB5E vector.Then the recombined phagemids were electroperated into E.coli TG1.The recombinant rate of scFv phage antibody library Was identified by enzyme digestion analysis and PCR.And the library repertoire Was determined by titration the PFU of library.Results The phage antibody library of scFv which specific binding to the proteins of H5N1 virus Was generated from the peripheral blood of the recovery phase patient infected with H5N1 virus and the repertoire of library was 3.75×104.Conclusion Phage antibody library of humanized scFv fragment which specific binding to the proteins of H5N1 virus has been constructed and screened successfully,which lay the foundation for further study of quick detection method and medicihal purpose. Key words: Human H5N1 virus; Humanized single chain antibody; Phage library; Construction; Screening