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Monoclonal antibody of equine influenza virus nucleoprotein as well as preparation method and application thereof

A monoclonal antibody, equine influenza technology, used in hybridoma cell lines, application of reagents, and the field of monoclonal antibodies, can solve the problems of limited application, difficult to popularize and use at the grassroots level, and difficulty in large-scale detection, and achieve good affinity. Effect

Inactive Publication Date: 2012-10-24
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the commonly used methods for detecting EI in China include virus isolation, hemagglutination inhibition test, RT-PCR, etc., but it is difficult to carry out large-scale detection, and the application is limited to professional laboratories, so it is difficult to promote the use at the grassroots level

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  • Monoclonal antibody of equine influenza virus nucleoprotein as well as preparation method and application thereof
  • Monoclonal antibody of equine influenza virus nucleoprotein as well as preparation method and application thereof
  • Monoclonal antibody of equine influenza virus nucleoprotein as well as preparation method and application thereof

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Embodiment Construction

[0015] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.

[0016] Embodiment Preparation and identification of monoclonal antibody of the present invention

[0017] 1. Materials and methods

[0018] 1.1 Virus strains, cells and experimental animals

[0019] EIV A / horse / Xinjiang / 07 (H3N8) was isolated and preserved by the inventor's laboratory, equine viral arteritis virus (EAV), equ...

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Abstract

The invention discloses a monoclonal antibody of an equine influenza virus (EIV) nucleoprotein (NP) as well as a preparation method and application thereof. The preparation method comprises the steps of: fusing cells with EIV NP expressed by pronucleus as immunogen, and then cloning the cells by utilizing a limiting dilution method and sifting the cells by utilizing an indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) to obtain two hybridoma strains 2G11 and 3E10 for stably secreting monoclonal antibodies. The two monoclonal antibodies can generate higher ascites titer. Western-blot analysis shows that the two obtained monoclonal antibodies can specifically identify EIV NP recombinant protein; an indirect immunofluorescence experiment proves that the two monoclonal antibodies can be combined with a natural EIV; and cross reaction is not carried out between the two monoclonal antibodies and equine herpesvirus type I, equine herpesvirus type IV and equine encephalitis virus type II through the ELISA. The invention provides the material basis for preparing an anti-EIV NP monoclonal antibody and establishing a subsequent EIV detection method.

Description

technical field [0001] The present invention relates to a monoclonal antibody, in particular to an equine influenza virus NP monoclonal antibody and a hybridoma cell line secreting the monoclonal antibody. The present invention also relates to the use of the equine influenza virus NP monoclonal antibody in the preparation of a reagent for detecting or diagnosing equine influenza virus The application in the present invention belongs to the detection field of equine influenza virus. Background technique [0002] Equine influenza (Equine influenza, EI) is an acute respiratory disease of equine animals caused by equine influenza virus (Equine influenza virus, EIV). Due to its long recovery period and high treatment costs, it has brought great economic benefits to the horse industry Loss (REEVE-JOHNSON L. Equine influenza in Australia [J]. Vet Rec, 2007, 161: 635.). Therefore, it is extremely important to establish a rapid and effective method for detecting EI for its early pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/569
Inventor 郭巍相文华赵立平
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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