Detection of HCV core protein,envelope glycoprotein and core-envelope fusion protein by baculovirus surface display |
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Authors: | LI Xiang JIN Ai-hui ZOU Li-lin GAO Ji-min L Jian-xin |
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Affiliation: | Zhejiang Provincial Key Laboratory of Medical Genetics, Zhejiang Provincial Key Laboratory for Technology and Application of Model Organisms, Wenzhou Medical College, Wenzhou 325035, China |
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Abstract: | AIM: To detect the hepatitis C virus (HCV) at early stages by the technique of baculovirus surface display. METHODS: We constructed the recombinant baculoviruses vAc-Flag-HCV C-GP64, vAc-Flag-HCV E-GP64 and vAc-Flag-CE-GP64, which displayed HCV-related structural proteins by the technique of baculovirus surface display. RESULTS: Western blotting results showed that the core (C), envelope (E) and core-envelope (CE) proteins of HCV were expressed in Sf9 cells after the infection of recombinant baculoviruses. We also observed by confocal microscopy that the C, E and CE proteins of HCV presented and were anchored on the plasma membrane of Sf9 cells after infection. In addition, the results of immunogold electron microscopy demonstrated that the 3 recombinant baculoviruses displayed the C, E and CE proteins of HCV on the viral surface, respectively. To further define the role of these recombinant baculoviruses in detecting HCV at early stages, we applied time-resolved fluoroimmunoassay (TRFIA) to detect HCV samples after purification and concentration of the recombinant baculoviruses. The results showed that the positive rate of vAc-Flag-CE-GP64 was up to 80.2%. CONCLUSION: vAc-Flag-CE-GP64 is useful for detecting the hepatitis C virus at early stages. |
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Keywords: | Hepatitis C virus Baculovirus surface display Time-resolved fluoroimmunoassay |
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