Construction of library of human lung cancer specific single chain antibodies via phage display

Objective:To construct a library of human lung cancer specific single chain Fv antibodies via phage display and establish the foundation for screening antibodies of lung cancer-associated antigen.Methods:Total RNA was extracted from(meta)static lymph node of lung cancer patients.The variable regions of heavy chain(VH) and light chain(VL) of human antibodies were amplified by RT-PCR.VH and VL were linked with each other to generate scFv gene in vitro.The scFv fragments were cloned into the phagemid(pCANTAB 5E) and the recombinant phagemid was transformed to susceptible E.coli TG1 by electroporation.They were superinfected by helper phages to form phage display library of single chain Fv antibodies. Finally,the diversity of the library was identified by restriction endonuclease digestion.Results:Total RNA was extracted successfully from lymph nodes of metastatic lung cancer patients.The variable regions of human antibodies was amplified by RT-PCR and(connected) with each other to form ScFv.The constructed phage display antibody library contained 1.2×10~8 independent clones.Result of BstN Ⅰdigestion showed that it had good diversity.Conclusion:The phage display library of human lung cancer specific single chain Fv antibodies is constructed successfully.It establishes the foundation for screening the soluble antibodies of lung cancer associated protein.