HCV核心蛋白片段的表达、纯化及免疫原性检测

目的：探讨HCV核心蛋白N端119aa片段表达产物的抗原性及用于抗-HCV抗体检测的可能性。方法：把HCV C119-pEq32a／BL21重组菌进行发酵表达，提取包涵体，并用阳离子柱进一步纯化目的蛋白。用Western blot及ELISA分析该融合蛋白的免疫特异性及敏感性。结果：所表达的目的蛋白约占菌体总蛋白的20％，SDS-PAGE结果表明所纯化的目的蛋白纯度大于95％。Western blot及ELISA结果显示该融合蛋白具有良好的免疫特异性，所检测的47份临床HCV感染血清的阳性率为91．5％，而阴性对照及HBV感染血清则未见阳性结果。结论：该HCV Cll9融合蛋白检测HCV感染患血清具有良好的特异性和敏感性，可望用于HCV抗体检测试剂盒的研制。

Expression, Purification and Antigenicity of a 119aa Fragment of the Core Protein of Hepatitis C Virus

Objective To analyze the sensitivity and specificity of a 119aa fragment of the core protein of hepatitis C virus in detection of anti HCV antibody. Methods The HCV C119 pET32a/BL21 recombinant bacteria were fermented and the fusion protein C119 were purified as a form of inclusion body and further purified by SP Sepharose HP column. Western blot and ELISA were used to analyze the antigenicity and immunospecificity of the C119 purified protein. Results The C119 protein was accounted for about 20% of total bacterial proteins and the purity of C119 was over 95% after two steps of purification. The results of Western blot and ELISA showed that C119 protein had excellent HCV immunospecificity and antigenicity. Using C119 pure recombinant for the detection of anti HCV antibody, 43 samples in all 47 HCV clinical positive sera showed positive while all the normal controls and HBV clinical negative showed negative.Conclusion The results indicated that the recombinant C119 protein demonstrated a high immunospecificity and antigenicity.It may be a useful agent in the detection of anti HCV antibody.