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Preparation of common antigen monoclonal antibody of Brucella sLPS (lipopolysaccharides) and establishment of c-ELISA (competitive enzyme-linked immuno sorbent assay)method

A monoclonal antibody, Brucella technology, applied in the field of immunology, can solve the problem that the chemical structure is not clearly identified, and achieve the effect of high application value

Inactive Publication Date: 2014-06-18
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

The chemical structure of the C epitope has not yet been clearly identified

Method used

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  • Preparation of common antigen monoclonal antibody of Brucella sLPS (lipopolysaccharides) and establishment of c-ELISA (competitive enzyme-linked immuno sorbent assay)method
  • Preparation of common antigen monoclonal antibody of Brucella sLPS (lipopolysaccharides) and establishment of c-ELISA (competitive enzyme-linked immuno sorbent assay)method
  • Preparation of common antigen monoclonal antibody of Brucella sLPS (lipopolysaccharides) and establishment of c-ELISA (competitive enzyme-linked immuno sorbent assay)method

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Experimental program
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Embodiment Construction

[0028] Hereinafter, the present invention will be described in detail with reference to Examples, which are only intended to illustrate the present invention and are not intended to limit the scope of the present invention. The scope of the invention is specifically defined by the appended claims.

[0029] 1 Materials and methods

[0030] 1.1 Main reagents

[0031] Proteinase K, DNase I, RNase, benzidine diamine (OPD), DMEM medium, and PEG3000 were all purchased from Sigma; HAT and HT selection medium were purchased from Gibco; the monoclonal antibody subclass identification kit was purchased from Southern Biotech Company; Protein G Sepharose column was purchased from GE Healthcare Company.

[0032] 1.2 Strains

[0033] Vaccine strains S-19, M5-90 and S2 Brucella, Escherichia coli (O: 157), Salmonella Dublin (C79-86), Yersinia enterocolitica (O: 9) and Scruff Rutella (M111) were all commercially purchased from China Veterinary Drug Control Institute.

[0034] 1.3 Experime...

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Abstract

The invention provides a monoclonal antibody against smooth Brucella (Brucella) lipopolysaccharides (sLPS), and preferably provides 14F4. Cross reaction tests show that the monoclonal antibody 14F4 has no cross reaction with Escherichia coli (O:157), Salmonella Dublin (C79-86) whole thallus and the LPS. On the basis, a competitive ELISA (c-ELISA) test method using the monoclonal antibody 14F4 as a competition antibody is established, and the method is suitable for testing large-scale clinical samples, and has the characteristics of being rapid, high-throughput, high-sensitivity and high-specificity.

Description

technical field [0001] The present invention relates to the field of immunology. Specifically, the present invention relates to a monoclonal antibody against the Brucella lipopolysaccharide common antigen, a c-ELISA method based on the monoclonal antibody, a kit containing the monoclonal antibody and a diagnostic detection reagent. Background technique [0002] Brucellosis is a zoonotic infectious disease caused by Brucella bacteria. Causes miscarriage, infertility and arthritis among other symptoms. People infected with Brucella produce more serious symptoms, such as wave fever, unbearable joint and muscle pain, loss of working ability, and also affect fertility, and severe cases can cause death. It is prevalent in more than 160 countries and regions. The brucellosis epidemic in my country was well controlled from the late 1980s to the early 1990s. Since 1993, the brucellosis epidemic in my country has been increasing year by year. The world suffers huge economic losses ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C12N5/20G01N33/577C12R1/91
Inventor 步志高胡森
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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