丙型肝炎病毒包膜蛋白E2抗原模拟表位的筛选和鉴定

Epitope mapping of hepatitis C virus E2 antigen from a 7 peptide phage library by using immobilized specific monoclonal antibody

目的 :筛选丙肝病毒 (HCV)包膜蛋白 (E2 )特异性噬菌体模拟表位。方法 :以抗 HCVE2的单克隆抗体作为固相筛选分子 ,对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 5 0个克隆 ,经酶联免疫吸附法 (ELISA)鉴定、交叉反应以及竞争抑制性实验 ,最后对所选克隆进行DNA序列分析 ,以确定HCVE2抗原的模拟表位。结果 :经噬菌体富集后 ,得到 12个阳性克隆 ,确定氨基酸序列XXPXYXW为HCVE2的模拟表位。结论 :用噬菌体七肽库成功筛选得到HCVE2的模拟表位 ,为开展用HCV模拟表位探索HCV的防治研究创造了条件。

Objective:To screen HCV E2 mimotopes by using monoclonal antibody and phage peptide library.Methods:By using HCV E2 monoclonal antibody as selective molecule, a 7 mer phage peptide library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing. Results:12 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparision results, one epitope was confirmed as mimotope of HCV E2. Conclusion: HCV E2 mimotope was obtained by phage peptide library screening. The result provides a new approach for HCV therapy and vaccine development.