A high-affinity anti-vomitoxin monoclonal antibody and its preparation method

A technology of monoclonal antibody and deoxynivalenol, which is applied in the field of immunoassay, can solve the problem of low affinity of anti-vomitin monoclonal antibody, and achieve the effects of overcoming low affinity, accurate detection, and cost reduction

Active Publication Date: 2018-01-19
SHANDONG LVDU BIO SICIENCE & TECH
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  • Application Information

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Problems solved by technology

[0003] At present, the detection methods of vomitoxin include chemiluminescence, liquid chromatography, enzyme-linked immunoassay, and immunocolloidal gold method. Among them, chemiluminescence and liquid chromatography require large-scale instruments and professionals, while other methods The core is to prepare high-affinity anti-vomitoxin monoclonal antibodies. At present, domestic anti-vomitoxin monoclonal antibodies have low affinity and are mostly imported.

Method used

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  • A high-affinity anti-vomitoxin monoclonal antibody and its preparation method
  • A high-affinity anti-vomitoxin monoclonal antibody and its preparation method

Examples

Experimental program
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Embodiment 1

[0026] Example 1 Preparation method of high-affinity anti-emetic toxin monoclonal antibody

[0027] (1) Deoxynivalenol complete antigen synthesis

[0028] Dissolve deoxynivalenol and N, N-carbonyldiimidazole (CDI) in pyridine at a mass volume ratio of 10:1 (mg:mL), and derivatize at 37°C for 3 hours to obtain the deoxynivalenol hapten. Add the hapten dropwise to Put into 0.01M PBS (3mL 0.01M PBS containing 20mg BSA) equivalent to 3 times the volume of pyridine, stir at room temperature for 1 hour, and dialyze with 0.01mol / LPBS for 2 days to obtain vomitoxin immune antigen. The synthesis process of vomitoxin complete antigen is as follows: figure 1 shown;

[0029] (2) Preparation of vomitoxin monoclonal antibody

[0030] ①Immunization program

[0031] The immune antigen obtained in step (1) was mixed with 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., product number LD003) at a ratio of 1:1, and the 8-10-week-old Balb / c mice (purchased from Beijing Weitongli China Exp...

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Abstract

The invention relates to a high-affinity anti-DON monoclonal antibody and a preparation method thereof. The preparation method includes the following steps: (1) complete antigen synthesis of DON; (2) preparation of DON monoclonal antibody. Step (2) includes immunization procedure, cell fusion, hybridoma screening and cloning, and antibody preparation and purification. The invention can quickly, sensitively and accurately detect the vomitoxin, and overcomes the problem that the domestic anti-vomitoxin monoclonal antibody has low affinity and is mostly imported.

Description

technical field [0001] The invention relates to a high-affinity anti-vomitoxin monoclonal antibody and a preparation method thereof, belonging to the technical field of immune detection. Background technique [0002] The toxin-producing bacteria of vomitoxin are mainly the dwarf fungus of the genus Fusarium, and vomitoxin mainly contaminates corn, wheat, rice, barley, millet and oats. Deoxynivalenol has an estrogen effect, mainly acting on the reproductive system, and can cause hyperestrogenism in livestock and poultry. Eating pasta made from vomitoxin-containing flour can cause central nervous system poisoning, with symptoms such as nausea, chills, headache, mental depression, and ataxia. Pregnant humans and animals eating food containing vomitoxin can easily cause miscarriage, stillbirth and teratogenicity. Therefore, it is of great significance to establish a rapid, sensitive and accurate detection technology for vomitoxin. [0003] At present, the detection methods of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/14G01N33/74G01N33/577C12N5/20
Inventor 武玉香沈志强王金良张莎莎于金枝
Owner SHANDONG LVDU BIO SICIENCE & TECH
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