人源性噬菌体抗体库的构建及HBsAg人单抗的筛选

CLONING OF HUMAN McAb AGAINST HBsAg FROM PHAGE ANTIBODY LIBRARY

用聚合酶链延伸反应(PCR)从人外周血淋巴细胞克隆出Fd段和к链基因,重组到表达载体pCOMB3中,构建了人源性噬菌体抗体库。将乙肝病毒表面抗原(HBsAg)固相化对噬菌体抗体库进行了三轮“吸附—洗脱—扩增”的筛选,使特异性噬菌体抗体得到了富集,筛选到了抗HBsAg的人单抗。

Immunoglobulin heavy chain Fd genes and K chain genes were amplified by RT-PCR from PBLs of a volunteer immunized with HBsAg vaccine. Phage antibody library was constructed with the Fd genes and K chain genes using pCOMB3 as vector. Three rounds selection against coated HBsAg showed specific enrichment of phage antibodies. After the third round selection 19 out of 24 clones exhibited HBsAg binding capacity. The specificity of the clones was verified by ELISAs and competition inhibition ELISA. The success of isolating human anti-HBsAg Fab proved the usefulness of phage display system in human McAb preparation.