Construction and screening of human phage display antibody library against lung adenocarcinoma

Abstract

Objective:To construct human single-chain variable fragment(ScFv)antibody library,and screen out antibodies against lung adenocarcinoma from the library.Methods:The total RNA was isolated from tumor adjacent lymph nodes of the lung adenocarcinoma patients and was used to amplify VH and VL genes by RT-PCR.VH and VL genes was joined with a DNA linker by SOE-PCR to form the ScFv.The gel purified ScFv gene repertoires were cloned into the phage vector pCANTAB5E to construct the primary phage library.Panning against lung adenocarcinoma cell line A549 was performed for four rounds and the phage library was identified.Results:A recombinant phage antibody library was successfully constructed.The fourth phage harvest yielded 115 times as much as that of the first one.During affinity screening,the antibody was enriched with the increase of panning rounds.Positive reactions to A549 were detected in 7 of 10 randomly selected clones,with a positive rate of 70%.Conclusion:A human phage-display antibody library has been successfully constructed.The selected ScFv fragment can specifically bind to human lung adenocarcinoma cell line A549.