用PCA方法选择HBV DNA多聚酶TP区VH抗体

Isolation of the variable fragments of heavy chain against the terminal protein region of hepatitis B virus polymerase with protein fragment complementation assay

目的:用PCA方法选择HBVDNA多聚酶TP区VH抗体.方法:从YeastDisplayscFvAntibodyLibrary中提取包含scFv抗体库的质粒pPNL6,扩增VH抗体片段.以HepG2.2.15细胞分泌的HBVDNA为模板,PCR扩增HBVDNA末端蛋白(TP).以TP区作为抗原,用PCA方法进行VH抗体选择.结果:PCR扩增的TP区共540bp.对HBVDNA多聚酶TP区进行抗体选择,用PCA方法从抗体库中选择出3个对应抗体,根据DNA序列和WernerMüller数据库可标明3个抗体.结论:对HBVDNA多聚酶TP区进行抗体选择,用PCA方法可以从抗体库中较为简便的选择出对应抗体.

AIM： To isolate the variable fragments of heavy chain （VH） against the terminal protein （TP） region of hepatitis B virus （HBV） polymerase （Pol） with protein fragment complementation assay （PCA）. METHODS： The plasmid pPNL6, which contained scFv antibody library, was extracted from Yeast Display scFv Antibody Library to amplify the VH antibody fragment. The HBV DNA secreted by HepG2.2.15 cells was used as the template to amplify HBV DNA TP. Then the VH antibodies were selected with PCA using the TP region as the antigen. RESULTS： After amplification, TP region with a length of 540 bp were obtained. Three TP antigen-specific VH antibodies were amplified by PCA, and then they were translated into amino acids according to the DNA sequences and Werner Müller databank. CONCLUSION： TP region antigen-specific VH antibodiescan be directly selected from HBV DNA Pol with PCA.