关键词: 噬菌体抗体库, 单链抗体, 肺腺癌

Abstract: Objective To construct a human phage single chain-antibody library, and sieve out the antibody ScFv against lung cancer from the library. Methods The total RNA was isolated from lymph node tissue of the lung cancer patients, and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined with a DNA linker by SOE-PCR to form the single chain variable fragment (ScFv) gene. ScFv gene was coloned into the phage vector pCANTAB5E. Panning against lung cancer cell line A549 were performed for four rounds. The positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased 115 times. Positive reactions to A549 were detected in 7 of 10 random clones, The positive rate was 70%. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusions ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has avidity specifically to human lung cancer cells.

Key words: phage antibody library, single chain variable fragment, lung adenocarcinoma