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Anti-dengue virus E protein monoclone antibody, preparation method and uses thereof

A monoclonal antibody, dengue virus technology, applied in antiviral immunoglobulins, antiviral agents, biochemical equipment and methods, etc., can solve the problems of loss of antigenic determinants, lack of natural structure and natural folding of recombinant proteins, etc. To achieve the effect of reducing cross-reaction or side effects

Inactive Publication Date: 2009-01-14
ARMY MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are still a large number of reports on the preparation of monoclonal antibodies using recombinant proteins of dengue virus (such as NS1, E protein), but due to the lack of natural structure and natural folding of recombinant proteins, important epitopes may be lost

Method used

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  • Anti-dengue virus E protein monoclone antibody, preparation method and uses thereof
  • Anti-dengue virus E protein monoclone antibody, preparation method and uses thereof
  • Anti-dengue virus E protein monoclone antibody, preparation method and uses thereof

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Embodiment Construction

[0042] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0043] 1. Preparation of monoclonal antibody against dengue virus E protein

[0044] (1) Virus purification

[0045] (A) Virus propagation: get Aedes albopictus C6 / 36 cells, use RPMI 1640 culture fluid, and the content percentage concentration is the fetal bovine serum of 10%, the glutamine that molar concentration is 2mmol / L, molar concentration is 10mmol / L of HEPES buffer, after culturing to a single layer, carry out the infection of type II dengue virus (DENV2) Tr1751 strain according to the multiplicity of infection (MOI)=1, after obvious cytopathic changes appear, collect the culture medium that contains virus Clear, centrifuge to remove cell debris, use VERO cells (African green monkey kidney cells) monolayer culture plaque method (Plaque assay) to measure the virus titer, adjust the virus titer to 5.0×10 6 PFU / ml, stored at -80°...

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Abstract

The invention discloses monoclonal antibody of anti-dengue virus E protein, which is generated through the secretion of the mice hybridoma cells 2B10 with the collection number of CGMCC No.2407; the monoclonal antibody provided by the invention has the advantages that the comprehensive effectiveness is high, the monoclonal antibody can be specifically combined with the dengue virus and can be applied to the preparation of the novel and effective medicine which can prevent and cure dengue virus infection and can also be manufactured into reagent kits for testing dengue virus.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a monoclonal antibody against dengue virus E protein, a preparation method and an application thereof. Background technique [0002] Dengue fever and dengue hemorrhagic fever / shock syndrome are acute infectious diseases transmitted by mosquitoes caused by dengue virus (Dengue virus), characterized by fever, rash, hemorrhage, shock, etc., with a high fatality rate. In recent years, dengue fever and The incidence of dengue hemorrhagic fever / shock syndrome is on the rise, and the World Health Organization has listed dengue hemorrhagic fever / shock syndrome, hepatitis, malaria, and tuberculosis as the most serious infectious diseases in the world. Dengue virus is a group B arbovirus, which belongs to Togaviridae and Flavivirus. The virus is composed of RNA genome and capsid protein. It is dumbbell-shaped, rod-shaped or spherical, with a diameter of 30-40nm. The mouse brain and tissue cel...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N5/18A61K39/42A61P31/14G01N33/577
CPCY02A50/30
Inventor 陈宗涛安静刘丽梅
Owner ARMY MEDICAL UNIV
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