应用细菌鞭毛递呈的随机肽库研究丙型肝炎病毒核心蛋白B细胞抗原表位

设计了利用大扬杆菌鞭毛蛋白递呈的随机十二肽库研究ＨＣＶ核心蛋白Ｂ细胞抗原位的实验程序：１利用大肠杆有达质粒ｐＱＥ－３０有达并纯化ＨＣＶ核心蛋白Ｐ１９；２利用Ｐ１９蛋白亲和层析纯化ＨＣＶ感染者血清的抗ＨＣＶ核心蛋白多克隆抗体；

EPITOPE MAPPING OF POLYCLONAL ANTIBODIES TO HCV CORE PROTEIN USING BACTERIAL RANDOM DODECAPEPTIDE LIBRARY

A novel method has been designed for epitope mapping of polyclonal antibodies to HCV core protein by bacterial random dodecapeptide library. The truncated core protein of HCV, P19, was expressed in E. coli and purified with Nickel chelate affinity chromatography. Using purified protein P19, the anti-core polyclonal antibodies in HCV-positive sera were purified by affinity chromatography. Using the purified antibodies as selective molecules, a bacterial random dodecapeptide library was biopanned, and positive clones were selected with ELISA and RIA. Amino acid sequences of displayed peptides were compared with the aa sequence of HCV core protein. Two linear epitopes were mapped at residues 17-29 and 31-42 of HCV core protein.