Anti-trionyx sinensis immune globulin monoclonal antibody and application thereof

A monoclonal antibody and immunoglobulin technology, applied in immunoglobulin, application, biochemical equipment and methods, etc., can solve the problems of research, detection and rapid diagnosis of aquatic pathogenic microorganisms, disease diagnosis and prevention Difficulties and other problems, to achieve the effect of broad application prospects

Inactive Publication Date: 2011-01-26
ZHEJIANG INST OF FRESH WATER FISHERIES
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Problems solved by technology

[0003] The present invention aims to solve the problem of using monoclonal antibodies to study the structure and function of aquatic immunoglobulins. There are very few monoclonal antibodies used in the research, detection and rapid diagnosis of aquatic pathogenic microorganisms, so that

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  • Anti-trionyx sinensis immune globulin monoclonal antibody and application thereof
  • Anti-trionyx sinensis immune globulin monoclonal antibody and application thereof

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Embodiment Construction

[0020] 1. Preparation of Chinese soft-shelled turtle IgM monoclonal antibody

[0021] (1) Preparation of soft-shelled soft-shelled turtle specific antigen: cut off the neck of Chinese soft-shelled turtle, collect its blood and place it in a glass bottle, and solidify it at room temperature. Place it at 37°C for 2 hours, then place it at 4°C for 10 hours, the serum is fully separated, centrifuged at 2000r / min to precipitate, and the supernatant is taken to make a SephadexG-200 chromatography column, the column length is 70cm, the diameter is 1.6cm, and the serum of Chinese soft-shelled turtle is treated with 50% (W / W) Ammonium sulfate aqueous solution was crudely extracted and concentrated by dialysis, then diluted with PBS (pH7.4), filtered through a 0.45 μm filter membrane, and loaded with PBS, the eluate was collected by an automatic sampler, and used The protein concentration of the eluate was tracked and measured by an ultraviolet spectrophotometer, and the first eluted pr...

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Abstract

The invention relates to an anti-trionyx sinensis immune globulin monoclonal antibody, preparing the materials according to the following steps: (1) trionyx sinensis specificity antigen: collecting trionyx sinensis disposed in a bottle, condensing at a room temperature, standing the trionyx sinensis for 1.5-3h at 35-38 DEG C, standing for 8-12h at 3-6 DEG C again, centrifugating, taking supernatant, preparing a chromatographic column, crudely extruding ammonium sulfate water solution and diluting after dialyzing and concentrating, loading after filtering and eluting, wherein the firstly eluted protein IgM is the trionyx sinensis specificity antigen; (2) polyclonal antibody: sampling blood of the antibody immune animal, separating antiserum and purifying to obtain the polyclonal antibody; (3) hybridoma cell line: fusing myeloma cell with the splenocyte for generating the anti-trionyx sinensis immune globulin monoclonal antibody to obtain the hybridoma cell line; and (4) monoclonal antibody of anti-trionyx sinensis immune globulin: generating the monoclonal antibody by the hybridoma cell line. The trionyx sinensis immune serum titer is detected by the monoclonal antibody. The invention is suitable for researching turtle etiology and immunology.

Description

technical field [0001] The invention belongs to anti-aquatic organism immunoglobulin monoclonal antibody and application thereof, in particular to anti-turtle immunoglobulin monoclonal antibody and application thereof. Background technique [0002] In recent years, with the development of soft-shelled turtle breeding industry, soft-shelled soft-shelled turtle diseases have become increasingly rampant. The occurrence and prevalence of fulminant diseases have hindered the healthy development of soft-shelled turtle breeding industry. To solve these problems, in addition to strengthening management and improving breeding technology, we must also pay attention to the research of its etiology, immunology and other fields, introduce immune prevention methods into the Chinese soft-shelled turtle breeding industry, and effectively enhance the active defense ability of Chinese soft-shelled turtles through immune means. Reduce the chance of disease. Based on this understanding and pr...

Claims

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Application Information

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IPC IPC(8): C07K16/42C12N15/06C12P21/08G01N33/577
Inventor 沈锦玉徐洋郝贵杰潘晓艺姚嘉赟尹文林
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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