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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 a specific phage mimotope of hepatitis C virus nonstructural protein NS5 (HCV NS5) and explore new ways to study anti-HCV vaccine. Methods The anti-HCV NS5 monoclonal antibody was used as the solid-phase screening molecule. The synthetic phage random heptapeptide library was subjected to 5 rounds of “adsorption-elution-amplification” screening process, and 30 clones were randomly selected. ELISA was used to identify and carry out cross-reaction experiments and competitive inhibition binding experiments. Finally, the selected clones were subjected to DNA sequence analysis to confirm the mimotope of HCV NS5 antigen. Results After phage enrichment, 12 positive clones were obtained from 30 randomly selected clones, and the mimotope of amino acid sequence QIRPTRQ was determined to be HCV NS5. Conclusion The mimic epitopes of HCV NS5 were successfully screened with phage heptapeptide library, which provided conditions for exploring the prevention and treatment of HCV infection with HCV mimotopes.