丙型肝炎病毒亚单位融合蛋白的表达及免疫原性测定

Expression of hepatitis C virus subunit fusion protein and analysis of its immunogenicity

目的获得高纯度的丙型肝炎亚单位融合蛋白并评价其免疫原性。方法利用原核表达系统，以pET—11d作为载体，在大肠埃希菌BL21（DE3）中经IPTG诱导表达获得融合蛋白，之后采用DEAE阴离子交换和镍离子柱亲和层析进行纯化。通过Western Blot验证融合蛋白的抗原活性。同时以该蛋白免疫实验动物后测定血清中的抗体滴度。结果成功获得很高纯度的丙型肝炎亚单位融合蛋白，EIA显示融合蛋白能够诱生出高滴度的抗体。结论原核表达系统表达的丙型肝炎病毒亚单位融合蛋白具有较强的免疫原性，为丙型肝炎病毒治疗性和预防性疫苗的研制提供了一条新的途径。

Objective Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity. Methods With the vector of pET-11d, fusion protein was expressed in Escherichia coli BL21 （DE3） after induced by IPTG. The protein was then purified by DEAE negative ion exchange chromatography and Ni^2＋ affinity chromatography. Western Blot analysis was used to detect the antigenicity of the fusion protein. At the same time, the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA. Results High purified hepatitis C virus subunit fusion protein was obtained successfully. The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers. Conclusion The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity. It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.