Monoclone antibody of H5 subtype avian influenza virus haemagglutinin protein, or its combination liveness fragment and uses thereof

A monoclonal antibody and avian influenza virus technology, applied in the direction of antibodies, viral peptides, anti-animal/human immunoglobulin, etc., can solve the problems that cannot meet the needs of clinical testing

Inactive Publication Date: 2008-07-16
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the existing technology is far from meeting the needs of clinical testing.

Method used

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  • Monoclone antibody of H5 subtype avian influenza virus haemagglutinin protein, or its combination liveness fragment and uses thereof
  • Monoclone antibody of H5 subtype avian influenza virus haemagglutinin protein, or its combination liveness fragment and uses thereof
  • Monoclone antibody of H5 subtype avian influenza virus haemagglutinin protein, or its combination liveness fragment and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0221] Embodiment 1: Preparation of anti-H5 subtype avian influenza virus HA gene monoclonal antibody

[0222] Antigen preparation:

[0223] The virus strain Ck / HK / Yu22 / 02 (H5N1) (referred to as Yu22) was used to inoculate 9-day-old fertilized chicken embryos, and after incubation at 30□ for 2 days, the chicken embryo liquid was collected to obtain the amplified Yu22 virus. Collect the live virus, inactivate it with 0.03% formalin formalin at 4°C, and detect the inactivated virus by HA to determine the titer of the inactivated virus solution (note: refer to the WHO operation for the specific methods of HA titer determination and HI detection guideline, we chose HA=1024, which was provided by the Department of Microbiology, University of Hong Kong).

[0224] mouse

[0225] Six-week-old female Balb / c mice were purchased from the Anticancer Center of Xiamen University and bred there for experiments.

[0226] Hybridoma preparation:

[0227] We use standard in vivo immunization...

Embodiment 2

[0242] Embodiment 2: Assembly of H5 subtype influenza virus HA antigen detection kit (enzyme-linked immunoassay, ELISA)

[0243] This kit uses the double-antibody sandwich method to detect the HA antigen of the H5 subtype influenza virus in the specimen. First, the monoclonal antibody against H5 influenza virus HA gene is pre-coated on the polyethylene microwell strip of the kit. When the cleaved H5 influenza virus HA antigen is added to the microwell, the pre-coated monoclonal antibody It can be captured, and then the enzyme-labeled monoclonal antibody added can also bind to it, and then the result can be judged by the degree of color development of the enzyme-catalyzed substrate. When the specimen does not contain influenza virus antigen or H5 influenza virus, the substrate will not develop color. Detectable specimens include excreta, oral and nasal secretions, whole virus or lysed virus cultured in chicken embryos, etc.

[0244] Preparation of microtiter plate:

[0245] ...

Embodiment 3

[0282] Embodiment 3: Assembly of H5 subtype influenza virus anti-HA antibody detection kit (enzyme-linked immunoassay, ELISA)

[0283] This kit uses a competitive method to detect H5 influenza virus HA antigen-specific antibodies in serum samples. Firstly, the monoclonal antibody against H5 subtype influenza virus HA is coated once on the microwell strip in the kit, and the recombinant expression antigen of H5 subtype influenza virus HA gene is coated twice. After adding the serum sample and the enzyme-labeled monoclonal antibody, the specific antibody in the sample competes with the enzyme-labeled monoclonal antibody to bind to the antigen on the enzyme-labeled plate. If the added serum sample can significantly inhibit the binding of the enzyme-labeled monoclonal antibody to the antigen , it indicates that the specimen contains specific antibodies to influenza virus H5 type HA antigen. When the specimen does not contain influenza virus antibody or H5 influenza virus antibody...

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Abstract

The invention provides a McAb that can be specifically binded with the HA protein of H5N1 avain influenza virus and a McAb that can prevent from at least 50 percent of McAb activity combining with the HA protein of H5N1 avain influenza virus, which can be used for detecting, diagnosing, preventing and treating the avain influenza virus, in particular to the H5N1 avain influenza virus. The invention also provides the relating hybridoma cell lines, the separated nucleic acid molecule and short peptide, as well as the drug combination, the medicine diagnosis equipment and the kit with the McAb included.

Description

[0001] related application [0001] The present invention application claims the priority right from the Chinese invention application 200610002312.1 submitted on January 26, 2006, and the entire content of this application is included in the present invention application. field of invention [0002] The present invention relates to a monoclonal antibody that can specifically bind to the hemagglutinin (HA) protein of H5 subtype avian influenza virus, its conservative variant or active fragment, or the relevant coding sequence of its polypeptide or polypeptide analogue, or the resulting A cell line of the above-mentioned monoclonal antibody, and a method for using the antibody or fragment for diagnosis and treatment. Background technique [0003] Since H5 avian influenza first broke out in geese on farms in Guangdong Province in my country in 1996 (Xu X et al., 1999, Virology), another strain of H5 virus derived from this virus has spread in poultry farms in Hong Kong ( April...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N5/12C12N15/13C12N15/63G01N33/53G01N33/569A61K39/395C07K14/11C12N15/44
Inventor 夏宁邵陈毅歆葛胜祥罗文新张军
Owner XIAMEN UNIV
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