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Anti-thiacloprid monoclonal antibody and application thereof

A monoclonal antibody and anti-thiacloprid technology, applied in instruments, analytical materials, peptides, etc., to achieve the effects of high accuracy, easy popularization and high sensitivity

Active Publication Date: 2015-10-14
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the monoclonal antibody of thiacloprid at home and abroad

Method used

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  • Anti-thiacloprid monoclonal antibody and application thereof
  • Anti-thiacloprid monoclonal antibody and application thereof
  • Anti-thiacloprid monoclonal antibody and application thereof

Examples

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Embodiment 1

[0017] Example 1: Preparation of anti-thiacloprid monoclonal antibody

[0018] 1. Immunization of mice:

[0019] (1) preparing the immunogen by coupling the immune hapten shown in formula (I) to BSA (bovine serum albumin);

[0020]

[0021] (2) Initial immunization: take healthy BALB / c female mice aged 6-8 weeks, prepare the prepared immunogen with PBS buffer to 1 mg / mL, mix it with an equal volume of Freund's complete adjuvant (FCA), and fully After emulsification, each mouse was intraperitoneally injected with 200 μL (i.e. 100 μg of immunogen);

[0022] (3) Booster immunization: three weeks after the first immunization, mix the immunogen (equal dose with the first immunization) and an equal volume of Freund's incomplete adjuvant (FIA), fully emulsify, and inject 200 μL intraperitoneally into each mouse Afterwards, booster immunization once every two weeks, a total of 4 immunizations; 3 days before cell fusion, the mice were injected with the immunogen, and the injection...

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Abstract

The invention relates to an anti-thiacloprid monoclonal antibody and an application thereof, and specifically relates to a monoclonal antibody specifically recognizing thiacloprid and an application of the monoclonal antibody. The invention relates to the field of biology. According to the invention, an artificial antigen is used for immunizing a BALB / c female mouse; myeloma cells SP2 / 0 are subjected to cell fusion with mouse spleen cells, such that hybridoma cells are prepared; through series screening and sub-cloning, a hybridoma cell strain stably secreting the anti-thiacloprid monoclonal antibody is obtained; the hybridoma cell strain is injected into a mouse through intra-abdominal injection, such that ascite fluid is prepared; the mouse ascite fluid is purified with an octanoic acid-ammonium sulfate method, such that the thiacloprid specific monoclonal antibody is prepared. The antibody does not cross react with other compounds. An enzyme-linked immuno-chromatographic analysis method established with the antibody can be used in detecting environment and agricultural product thiacloprid residue rapidly, sensitively and conveniently with a low cost.

Description

technical field [0001] The invention relates to an anti-thiacloprid monoclonal antibody, especially a monoclonal antibody capable of specifically recognizing thiacloprid, and the application of the monoclonal antibody, belonging to the field of biotechnology. Background technique [0002] Thiacloprid, chemical name (3-((6-chloro-3-pyridyl)methyl)-1,3-thiazoline-2-ylidene) cyanamide, is a new type of nicotine chloride Insecticides, developed by Bayer Agrochemicals, have specific effects on piercing-sucking and chewing mouthparts pests. The mechanism of action is different from other traditional insecticides. By binding to nicotinic acetylcholine receptors, it interferes with the normal conduction of the insect nervous system, causing blockage of nerve channels, resulting in a large accumulation of acetylcholine, which makes the insects extremely excited, and the whole body is convulsed and paralyzed. and die. With strong contact, stomach poisoning and systemic effects, thia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44G01N33/577
Inventor 王鸣华华修德殷维施海燕
Owner NANJING AGRICULTURAL UNIVERSITY
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