Screening and identification of neutralizated single-chain antibody of anti-glomerular basement membrane antibody

VernacularTitle:中和抗肾小球基底膜抗体的单链抗体的筛选与鉴定
Author: Jing XIAO ; Zhangsuo LIU ; Pei WANG ; Liuyu HUANG ; Hongbin SONG ; Minghui ZHAO
Publication Type:Journal Article
Keywords: Phage display antibody library; Anti-glomerular basement membrane antibody
From: Chinese Journal of Microbiology and Immunology 2010;30(9):791-794
CountryChina
Language:Chinese
Abstract: Objective To screen a human single-chain variable fragments(scFv)against antiGBM antibody.Methods Using phage display technique,the phage antibody library was panned by antiglomerular basement membrane(GBM)antibody which was coated in a micro-titer plate,one clone was found to have high affinity to anti-GBM antibody.The DNA sequence of the positive clone was determined.Results Along with the increase of rounds anti-GBM antibody specific phage antibody was highly enriched and screening efficiency was increased 137 folds than the firest round.ELISA and competition inhibition assay showed that the scFv had a specific combination character with anti-GBM antibody.DNA sequencing confirmed that the whole gene of scFv was 750 bp,and in accordance with humanized single-chain variable region antibody sequence structure.Conclusion The results suggested that the scFv fragment to anti-GBM antibody could be successfully selected by recombinant phage antibody technique,which will laid an experimental foundation for further research of the therapy of Goodpasture syndrome.