HCV包膜蛋白E2的B细胞抗原模拟表位的筛选、重组表达与抗原性鉴定

Selection,Recombinant Expression and Antigenic Identification of an Mimotope of Hepatitis C Virus Envelope Protein E2

目的 采用蛋白质 /多肽递呈技术对HCV包膜蛋白E2进行模拟表位研究。方法 通过使用E2蛋白结合血清中的抗 E2 ,筛选M1 3噬菌体构建的随机肽库 ,寻找E2区蛋白的模拟表位 ;用大肠杆菌硫氧还蛋白作为骨架 ,在其活性部位以“内融合”形式重组表达该模拟表位 ,用HCV感染者血清验证重组蛋白的抗原性。结果 寻找到一个E2区蛋白的模拟表位 ,可能为E2蛋白的构象性表位。结论 本实验为研究HCV外膜蛋白E2抗原表位水平的特异诊断试剂的研制。

Objective To study the mimotoes of hepatitis C virus,envelope protein E2.by a biopanned phage library display ing random dodecamers.Methods Protein E2 was expressed and purified with pQE30 in M15.The purified protein E2 was used to bind anti E2 antibodies.After three rounds of screening,the positive phages were tested for their reactivity with antigen antibody of E2,P16 and HCV( ) IgG antibodies.Phages that showed positive reactivity with antigen antibody of E2,but negative to P16 and HCV( ) IgG antibodies,were selected and their displayed peptides were determined by DNA sequencing.Results One mimotope was selected to demonstrate its reactivity to anti E2 antibodies.Conclusion Through the expression of this small peptide in pTrxFus Expression System,the mimotope was demonstrated to be a conformational epitope of protein E2.