丙型肝炎病毒非结构蛋白NS4A抗原模拟表位的筛选和鉴定

Epitope mapping of hepatitis C virus NS4A antigen from a peptide phage library by using immobilized specific monoclonal antibody

目的 筛选丙型肝炎病毒 (hepatitisCvirus,HCV)非结构蛋白NS4A(HCVNS4A)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 HCVNS4A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 1 2肽库进行 5轮“吸附 洗脱 扩增”的筛选过程 ,随机挑取 45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HCVNS4A抗原的模拟表位。结果 经噬菌体富集后 ,从随机筛选的 45个克隆中得到 1 3个阳性克隆 ,确定氨基酸序列XXRXXMXPXXXI为HCVNS4A的模拟表位。结论 用噬菌体 1 2肽库成功筛选得到HCVNS4A的模拟表位 。

Objective To screen HCV NS4A mimotopes by using monoclonal antibody and phage peptide library.Methods By using HCV NS4A monoclonal antibody as selective molecule, a 12 mer phage peptide library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing.Results 13 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was confirmed as mimotope of HCV NS4A. Conclusion HCV NS4A mimotope was obtained by phage peptide library screening. The result provides a new approach for HCV therapy and vaccine development.