Coding region of hepatitis C virus in different His fusion expression and purification of antigen.docVIP

 PAGE \* MERGEFORMAT 26 Coding region of hepatitis C virus in different His fusion expression and purification of antigen Author: HAN cell, Qiao Wei Zhen, ZHANG Min, Sun Yanwen, Dong Chen, Ji-Hong Meng 【Abstract】 Objective: Expression and purification of hepatitis C virus (HCV)  Core, NS3 and NS4 of His fusion antigen H  C, H  NS3 and H  NS4, and to explore its application in antibody detection. Methods: Using recombinant genetic engineering techniques to construct three kinds of recombinant plasmid C  pET  28a  c (), NS3  pET  28a  c (), and NS4  pET  28a  c () and the corresponding engineered bacteria; quality tablets by the double digestion, PCR and sequencing is correct, IPTG-induced antigen expression, from the use of IPTG concentration, induction temperature and time three ways to optimize the antigen expression conditions; with NTA column purified antigens, respectively, a single recombinant antigen fragments and hybrid antigen by enzyme-linked immunosorbent assay (ELISA) detection of 100 HCV antibody-positive sera and 40 healthy human serum. Results: The single segment recombinant antigen and mixed antigen of 100 HCV antibody-positive serum, H  C, H  NS3 and H  NS4 antibody positive rates were 91%, 75% and 52%, while the H  C , H  NS3 and mixed antigen H  NS4 antibody detection rate was 100%; testing 40 healthy human serum, and all the results were negative. Conclusion: HCV coding region of a single fragment of different His fusion antigen with a different antibody detection rate, but lower than mixed-antigen positive rate; HCV antibodies in the development of diagnostic reagents should be mixed with HCV antigens. 【Key Words】 Hepatitis C virus; His fusion antigen; antigen purification; expression of HCV infection commonly used diagnostic method is mainly used ELISA for detection of anti  HCV. At present, HCV antibody detection kit at home and abroad have developed a three-generation product, the coating a