不同HCV优势抗原嵌合基因在大肠杆菌中的表达及其产物的分析

Expression and characterization of different chimeric antigens of hepatitis C virus

目的：为满足丙型肝炎病毒（ＨＣＶ）ＥＩＡ检测的需要，构建含优势抗原表位的ＨＣＶ嵌合重组抗原。方法：利用基因工程重组技术，获得了不同的ＨＣＶ嵌合抗原ＮＣ１和ＮＣ２。含嵌合基因的表达质粒在大肠杆菌中表达了目的抗原，其中包括结构区核壳蛋白Ｃ２２和不同长度的非结构区蛋白ＮＳ３；表达产物经离子交换和盐析纯化。结果：用针对Ｃ２２或ＮＳ３抗原表位特异性血清及系列血清进行了检测，表明ＮＣ１和ＮＣ２均同时具有Ｃ２２和ＮＳ３抗原的免疫反应性。以嵌合抗原制备的抗ＨＣＶＥＩＡ试剂检测中国药品生物制品检定所的第三代Ｐａｎｅｌ血清时，其阳性、阴性通过率都符合要求。结论：构建的嵌合抗原有多个优势抗原表位。

Objective: To obtain recombinant proteins which will be suitable for preparing diagnostic reagents and using in HCV immunology research. Methods: Recombinant antigens NC1 and NC2, possessing putative HCV nucleocapsid protein(C22) and different nonstructural protein 3(NS3)epitopes, were heavily expressed in E.coli, and the fusion proteins were then purified. Results: The purified recombinant antigens strongly react with sera containing human antibodies directed against C22 and NS3. The chimeric antigens obtained showed high sensitivity, specificity and good accordance with other commercial antiHCV ELISA kits when they were used as capture antigen in anti-HCV ELISA kit. Conclusion: The recombinant chimeric antigens can be good reagents for anti-HCV ELISA kit.