Anti-glutamate dehydrogenase protein monoclonal antibody, preparation method thereof and use thereof

A technology of glutamic acid dehydrogenase protein and monoclonal antibody, which is applied in the direction of anti-enzyme immunoglobulin, biochemical equipment and methods, antibodies, etc., can solve the problems of inability to meet clinical needs, lack of effective means for diagnosis and prevention, and virulence The problem of complex and unequal factors

Inactive Publication Date: 2010-08-18
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

However, due to the variety of bacteria types and complex virulence factors, and the pathogenic mechanism, interaction and relationship of each virulence factor are still unclear, there is still a lack of effective means for the diagnosis and prevention of the disease. The method still cannot meet the clinical needs, so how to establish a fast and accurate typing method needs to be further explored

Method used

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  • Anti-glutamate dehydrogenase protein monoclonal antibody, preparation method thereof and use thereof
  • Anti-glutamate dehydrogenase protein monoclonal antibody, preparation method thereof and use thereof
  • Anti-glutamate dehydrogenase protein monoclonal antibody, preparation method thereof and use thereof

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Embodiment 1

[0022] 1 Materials and methods

[0023] 1.1 Materials, experimental animals and reagents

[0024] Recipient Bacteria BL 21 , pET-30a, SP2 / 0 cells were preserved by our laboratory; BALB / c mice for experiments were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; Streptococcus suis standard bacteria SS1, SS2, SS7, SS9 were purchased from Harbin Veterinary Preserved in the institute, Freund's adjuvant, selection reagents HAT, HT, and fusion agent PEG were purchased from Sigma; standard fetal bovine serum and culture medium RPMI1640 were purchased from Gibco; restriction enzymes Sal I, BamH I, IPTG, antibiotics purchased from Dalian Bao Biological Engineering Co., Ltd.

[0025]1.2 Isolation and identification of WC-SS7 strain and preparation of genomic template DNA

[0026] In November 2007, a sick pig was sent to the laboratory in Wuchang City, Heilongjiang Province. After the autopsy, the brain an...

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Abstract

The invention discloses an anti-glutamate dehydrogenase protein monoclonal antibody, a preparation method thereof and use thereof. In the invention, the antigenicity and hydrophobicity of the GDH gene of streptococci are analyzed, a pair of primers, after a signal peptide and a transmembrane domain are removed, is designed to clone the GDH gene, a GDH protein of WC-SS7 is successfully expressed by Escherichia coli, and a BALB / c mouse is immunized with the purified GDH recombinant protein to obtain a hybridoma cell line (with a microorganism preservation number: CGMCC No.3343) which can react with the GDH protein and stably secrete the antibody. The obtained GDH recombinant protein and the specific monoclonal can be used in researches on the structure and functions of Streptococcus suis, so a foundation is built for the method for serodiagnosis of clinic infection with Streptococcus suis. The antibody, the preparation method, and the use have great values in aspects such as further research on the function of the glutamate dehydrogenase in the pathogenic mechanism of Streptococcus suis and the establishment of a Streptococcus suis detection method.

Description

technical field [0001] The present invention relates to a monoclonal antibody, in particular to an anti-glutamate dehydrogenase (GDH) protein monoclonal antibody and a preparation method thereof. The present invention also relates to the application of an anti-GDH protein monoclonal antibody in diagnosis or prevention and treatment of Streptococcus suis, belonging to The field of diagnosis or prevention of swine streptococcosis. Background technique [0002] Streptococcus suis is an important zoonotic pathogen. Pigs are generally susceptible to this disease. After infection, there may be acute sepsis, pneumonia, meningitis, arthritis, endocarditis and acute death. A variety of clinical types, if not treated in time, the mortality rate can reach 40% or even higher; the pigs that have endured grow slowly, and the storage time is greatly prolonged. Human infection with the bacteria can lead to meningitis, permanent deafness, sepsis, endocarditis, and even death. There are man...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/40G01N33/577A61K39/40A61P31/04C12R1/91
Inventor 王淑杰蔡雪辉徐敏武佳斌刘永刚石文达李丽琴徐明明荣福龙
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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