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Preparation method of benzothiostrobin hapten, artificial antigen and specific antibody and application thereof

A technology of benthiazolin and antibody heavy chain, applied in chemical instruments and methods, specific peptides, animal/human proteins, etc., to achieve the effects of easy popularization, low analysis cost, and large analysis capacity

Active Publication Date: 2015-05-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no reports about the artificial haptens, artificial antigens and antibodies of penthiostrobin at home and abroad

Method used

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  • Preparation method of benzothiostrobin hapten, artificial antigen and specific antibody and application thereof
  • Preparation method of benzothiostrobin hapten, artificial antigen and specific antibody and application thereof
  • Preparation method of benzothiostrobin hapten, artificial antigen and specific antibody and application thereof

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Experimental program
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Embodiment Construction

[0022] 1 Synthesis of artificial hapten

[0023] Alkaline hydrolysis of the carboxylate on the benzene ring of penthiostrobin to synthesize a hapten with a three-atom linking arm, which not only retains the characteristic group of penthiostrobin to the greatest extent, but also forms a carboxyl terminal group, which can Better for coupling to proteins. The product was purified by mass spectrometry (ESI) and H NMR spectroscopy ( 1 H-NMR) identification. The molecular structural formula of the artificial hapten of benzathystrobin is:

[0024]

[0025] 1.1 Synthesis of artificial haptens

[0026] Weigh 8.02g (0.02mol) of penthiostrobin technical substance and 30mL of tetrahydrofuran into a 100mL conical flask, then add 10.08mL of 5% lithium hydroxide solution (penthiostrobin:lithium hydroxide=1:1.05), Stir the reaction at room temperature for 4 hours, add 20 mL of water, extract three times with ethyl acetate, 40 mL each time, discard the organic phase, adjust the pH value...

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Abstract

The invention relates to a preparation method of a benzothiostrobin antigen and antibody and an application thereof, belonging to the technical field of immunochemical analysis. The chemical name of benzothiostrobin is 2-[[(5-methoxy-2-benzothiazole)-thiomethyl]-alpha-(E)-methoxymethylene]methyl phenylacetate. Under an alkaline condition, carboxylic ester in the benzothiostrobin structure is hydrolyzed to synthesize an artificial hapten of which the chemical name is 2-[[(5-methoxy-2-benzothiazole)-thiomethyl]-alpha-(E)-methoxymethylene]phenylacetic acid; the artificial hapten is coupled with bovine serum albumin and ovalbumin respectively to prepared an artificial antigen. The BALB / c female mouse is immunized by the artificial antigen to obtain a specific monoclonal antibody of benzothiostrobin. The antibody does not experience a cross reaction with other compounds. An enzyme linked immunosorbent assay method established by use of the antibody can be applied to quick, sensitive, convenient and cheap detection of benzothiostrobin residues in environment and agricultural products.

Description

1. Technical field [0001] The present invention relates to a preparation method of benzathystrobin artificial hapten, artificial antigen and specific antibody and its application. 2. Background technology [0002] Strobilurins fungicides are a new type of fungicides successfully developed under the guidance of natural antibiotics. They have the characteristics of broad spectrum, high efficiency and environmental friendliness. Benzothiostrobin is a new type of Strobilurins fungicide developed by Central China Normal University. Penthiostrobin has the advantages of wide bactericidal spectrum, high bactericidal activity, low application rate, long-lasting effect, etc. It has protective and therapeutic effects, and has excellent control effects on crop powdery mildew and downy mildew, and has a good market Development prospects. In order to prevent the potential risks in the application of penthiostrobin, a sensitive, rapid and selective residue detection method is needed. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D277/74C07K14/765C07K14/77C07K16/44G01N33/577
CPCC07D277/74C07K14/765C07K14/77C07K16/44C07K19/00G01N33/577
Inventor 王鸣华袁玉龙华修德施海燕
Owner NANJING AGRICULTURAL UNIVERSITY
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