Javadi Foozieh, Rahimi Pooneh, Modarressi Mohammad Hossien, Bolhassani Azam, Ardestani Mehdi Shafiee, Sadat Seyed Mehdi, Aghasadeghi Mohammad Reza
Clin Lab. 2016 Jul 1;62(7):1271-1278. doi: 10.7754/Clin.Lab.2015.151118.
Hepatitis C virus infection is one of global health concern. No vaccine is available so far and nonstructural protein 3 (NS3) is one of the main target antigens for developing studies on therapeutic vaccine and diagnostic application. In the current study, we expressed a truncated recombinant HCV-NS3 protein under native condition and evaluated its potential applications in immunization and diagnosis.
The recombinant pET-NS3 containing a truncated form of HCV NS3 region was constructed, confirmed by sequencing reactions, and expressed into E.coli BL21-DE3 strain. The expressed protein was purified by affinity chromatography under native condition. Characterization of diagnostic value and immunogenicity of this recombinant protein were analyzed by using an indirect ELISA method.
Our data showed that a truncated NS3 which contains the immunodominant region of this protein was successfully expressed and purified with a high yield of 3 mg/L. Our data showed that the immunogenicity of this truncated protein can induce a specific humoral response, as well as the usage for screening of HCV positive blood samples.
Altogether, the present study provides a simple and efficient system for protein expression and purification of an immunodominant region of NS3-HCV in native conformation and its potential application for diagnosis and vaccine development in future.
丙型肝炎病毒感染是全球关注的健康问题之一。目前尚无疫苗可用，非结构蛋白3（NS3）是治疗性疫苗和诊断应用研究的主要靶抗原之一。在本研究中，我们在天然条件下表达了一种截短的重组HCV-NS3蛋白，并评估了其在免疫和诊断中的潜在应用。
构建了含有截短形式HCV NS3区域的重组pET-NS3，通过测序反应进行确认，并在大肠杆菌BL21-DE3菌株中表达。表达的蛋白在天然条件下通过亲和层析进行纯化。使用间接ELISA方法分析该重组蛋白的诊断价值和免疫原性。
我们的数据表明，含有该蛋白免疫显性区域的截短NS3成功表达并纯化，产量高达3mg/L。我们的数据表明，该截短蛋白的免疫原性可诱导特异性体液反应，以及用于筛选HCV阳性血样。
总之，本研究提供了一个简单有效的系统，用于天然构象的NS3-HCV免疫显性区域的蛋白表达和纯化，及其在未来诊断和疫苗开发中的潜在应用。
