丙型肝炎病毒非结构蛋白NS_5抗原模拟表位的筛选和鉴定

Epitope mapping of hepatitis C virus non-structure protein 5 from a 7 peptide phage library by using immobilized specific monoclonal antibody

目的 筛选丙型肝炎病毒非结构蛋白NS5（HCV NS5）特异性噬菌体模拟表位,为抗HCV的疫苗研究探索新途径。方法 以抗-HCV NS5的单克隆抗体作为固相筛选分子,对人工合成的噬菌体随机七肽库进行5轮“吸附-洗脱-扩增”的筛选过程,随机挑取30个克隆,经噬菌体酶联免疫吸附法（ELISA）鉴定并进行交叉反应实验以及竞争抑制性结合实验,最后对所选克隆进行DNA序列分析,以确定HCV NS5抗原的模拟表位。结果 经噬菌体富集后,从随机筛选的30个克隆中得到12个阳性克隆,确定氨基酸序列QIRPTRQ为HCV NS5的模拟表位。结论 用噬菌体七肽库成功筛选得到HCV NS5的模拟表位,为用HCV模拟表位探索HCV感染的防治研究创造了条件。

Objective To screen HCV NS5 mimotopes by using monoclonal antibody and phage peptide library. Methods By using HCV NS5 monoclonal antibody as selective molecule, a 7 peptide phage library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing. Results Twelve positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was confirmed as the mimotope of HCV NS5. Conclusions HCV mimotope is obtained by phage peptide library screening. The result provides a new approach for HCV therapy and vaccine development.