Preparation and application of a novel HCV diagnostic antigen fused to streptavidin
10.3760/cma.j.issn.1003-9279.2011.03.023
- VernacularTitle:HCV免疫学诊断抗原与链亲和素融合蛋白的制备及应用研究
- Author:
Tingying ZHANG
1
;
Jian LU
;
Min ZHAO
;
Guo-Xia ZHAO
;
Yao DENG
;
Sheng-Li BI
;
Ji-Min GAO
;
Wen-Jie TAN
Author Information
1. 温州医学院
- Keywords:
Hepatovirus;
Antigens;
Streptavidin;
Viral fusion proteins
- From:
Chinese Journal of Experimental and Clinical Virology
2011;25(3):230-232
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare streptavidin-tagged hepatitis C virus (HCV) fusion protein and explore its application for the detection of antibody against HCV infection. Methods A recombinant plasmid pET-11 d-C44P-SA was constructed, which coding a novel HCV diagnostic antigens( C44P) and streptavidin (SA) fusion protein, and the fusion protein was generated with BL21(DE3) E Coli and identified by Western Blot analysis. Then the fusion protein was purified through the Ni-NTA affinity chromatography and over 90% purity has been achieved. Anti-HCV ELISAs were developed when the fusion protein was used in the biotin-pre-coated microplate or ordinary microplate, and then the sensitivity and specificity of the ELISA were evaluated with confirmed human sera panels. Results The fusion protein was expressed in high yields and purified successfully, the ELISA detection of anti-HCV with human sera panel indicated that its sensitivity and specificity is higher when SA-tagged HCV antigen ( C44P-SA) coated in biotin-pre-coated microplate, compared to C44P or C44P-SA coated in ordinary microplate. Conclusion The sensitivity and specificity of anti-HCV ELISA can be improved when a novel HCV diagnostic antigen fused to SA combined with the biotin- pre-coated microplate. This study laid a foundation for improving the performance of HCV diagnostics.
