Construction of a large human natural phage single-chain antibody library

Objective: To construct a human natural phage single -chain antibody (scFv) library with diversity.Methods: Total RNA was extracted from the peripheral blood lymphocytes of the non-immunized healthy donors, VH and VL genes were amplified by RT-PCR and were assembled to form scFv by overlap PCR and cloned into phagemid pCANTAB5E, and then transformed into E. coli TG1 by electroporation to construct a human natural phage scFv antibody library. Results:VH gene families and VL gene families were successfully amplified. A large human antibody library containing 6×108 clones was created after rescuing the recombinant phagemids from the transformed E. coli TG1 cells.Conclusion:A human scFv antibody library is successfully constructed finally. In the future, we can try to obtain antibody by panning purposely using related antigens.