Expression of Hepatitis C Virus Core and E2 antigenic recombinant proteins and their use for development of diagnostic assays

International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases
Amjad AliMuhammad Iqbal

Abstract

Early diagnosis of HCV infection is based on detection of antibodies against HCV proteins using recombinant viral antigens. The present study was designed to select, clone and express the antigenic regions of Core and E2 genes from local HCV-3a genotype and to utilize the antigenic recombinant proteins (Core & E2) to develop highly sensitive, specific and economical diagnostic assays for detection of HCV infection. The antigenic sites were determined within Core and E2 genes and were then cloned in pET-28a expression vector. The right orientation of the desired inserted fragments of Core and E2 were confirmed via sequencing prior to expression and were then transformed in BL21 (DE3) pLysS strains of E. coli and induced with 0.5mM Isopropyl-b-D-thiogalactopyranoside (IPTG) for the production of antigenic recombinant proteins. The produced truncated antigens were then purified by Nickel affinity chromatography and were confirmed by western blotting, immunoblotting and enzyme-linked immunosorbent assay (ELISA). The expressed Core and E2 recombinant antigens were used to develop immunoblotting assay for the detection of anti-HCV antibodies in sera. With immunoblotting, a total of 93-HCV infected sera and 35-HCV negative individuals...Continue Reading

References

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Sep 26, 1997·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·A S Lok, N T Gunaratnam
Aug 23, 2003·Seminars in Liver Disease·Jean-Michel Pawlotsky
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Sep 11, 2007·Advanced Drug Delivery Reviews·Tetsuro SuzukiTakaji Wakita
Sep 11, 2012·International Journal of Molecular Medicine·Tingxiu XiangGuoqi Lai

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