摘要
目的:筛选丙型肝炎病毒(HCV)非结构蛋白NS5A(HCVNS5A)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:应用噬菌体表面展示技术,以抗-HCVNS5A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12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HCVNS5A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XXXPXXXLLRXX为HCVNS5A的模拟表位.结论:用噬菌体12肽库成功筛选得到HCVNS5A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.
AIM:To screen HCV NS5A mimotopes by using monoclonal antibody and phage peptide library. METHODS:By using HCV NS5A monoclonal antibody as selective molec ule, a 12 mer phage peptide library was biopanned and positive clones were selec ted by ELISA, competition assay and DNA sequencing. RESULTS:12 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was confirmed as mimotope of HCV NS5A. CONCLUSION:HCV NS5A mimotope was obtained by phage peptide libra ry screening. The result provides a new approach for HCV therapy and vaccine de velopment.
出处
《世界华人消化杂志》
CAS
2002年第2期133-136,共4页
World Chinese Journal of Digestology
基金
国家自然科基金资助项目
No.39900130~~
