Preparation and application of chimeric VLPs (virus-like particles) expressing multiple series-connected HCV (hepatitis C virus) neutralization antigenic epitopes
A technology of antigenic epitopes and chimeric viruses, applied in the fields of molecular biology and cell biology, can solve problems affecting the packaging of virus particles, and achieve the effect of simple preparation and easy concentration and purification
Inactive Publication Date: 2016-10-12
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology
The hydrophilic region of HBV S antigen can insert epitopes, and express the conserved epitopes of pathogens. However, studies have found that the insertion of epitopes in the hydrophilic region is longer than 36 amino acids, which will affect the packaging of virus particles.
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[0036] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.
[0037] Using the method of chimerizing the target gene into the N-terminal extracellular region of HBV S without changing its transmembrane domain (TMD), the HCV neutralizing antigen epitope is chimerized at the N-terminal of the HBV S antigen in series to construct genotype C Hepatitis virus tandem multi-neutralizing antigen epitopes and hepatitis B virus S antigen chimeric gene eukaryotic expression plasmid, observed its expression in 293T cells, and prepared chimeric HCV tandem multi-neutralizing antigen epitopes HBV S antigen VLPs.
[0038] 1. Gene selection and synthesis of tandem hepatitis C virus multi-neutralizing antigen epitopes
[0039] 1.1 Epitope selection includes:
[0040] HCV E1 linear neutralizing epitope: ITGHRMAWDMMMNWS;
[0041] HCV E2 linear neutralizing epitope: QLINTN...
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The invention discloses a preparation method and an application of chimeric VLPs (virus-like particles) expressing multiple series-connected HCV (hepatitis C virus) neutralization antigenic epitopes and belongs to the fields of molecular biology and cytobiology. According to the method, a target gene is subjected to chimerism to an amino-terminal extracellular region of HBV S (hepatitis B virus S) without changing a TMD (transmembrane domain) mode, the HCV neutralization antigenic epitopes are connected in series and then subjected to chimerism to HBV S antigen amino terminals, eukaryotic plasmids expressing multiple series-connected HCV neutralization antigenic epitopes and HBV S antigen chimeric genes are constructed, expression of the plasmids in 293T cells are observed, and the VLPs of the HBV S antigen with multiple chimeric series-connected HCV neutralization antigenic epitopes are prepared. The VLP immune serum has certain protection effect on in-vitro simulated HCV infection, so that a new technical method is provided for design of an HCV neutralizing antibody vaccine, and thought is provided for design of a bivalent vaccine.
Description
technical field [0001] The invention belongs to the fields of molecular biology and cell biology, and specifically relates to a tandem HCV multi-neutralizing antigen epitope chimeric virus-like particle and a preparation method and application thereof. Background technique [0002] Chronic liver disease caused by HCV infection is a global public health problem, and the number of deaths caused by HCV infection reaches 450,000 worldwide every year. Due to the long-term lack of suitable in vitro virus culture systems, small animal models and accurate neutralizing antibody evaluation methods, the development of HCV preventive vaccines has not been successful so far. Studies have shown that there are multiple conserved neutralizing antigenic epitopes in the E1 / E2 region of the HCV envelope protein, and monoclonal antibodies against these epitopes have broad-spectrum cross-neutralizing activities, which is also the basis for designing the prevention of neutralizing antibodies Sex...
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Login to View More IPC IPC(8): C12N7/04C12N15/85C07K14/18A61K39/295A61K39/29A61P31/14
CPCC12N7/00A61K39/12A61K2039/5258A61K2039/55566A61K2039/70C07K14/005C12N15/85C12N2770/24223C12N2770/24234
Inventor 韦三华王晓岩张海雷迎峰林芳崔颖李斌张惠中
Owner FOURTH MILITARY MEDICAL UNIVERSITY