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Pyrethroid pesticide artificial antigen, antibody and preparation thereof

A pyrethroid, artificial antigen technology, applied in chemical instruments and methods, animal/human proteins, serum albumin and other directions, can solve the problems of high preparation cost, many synthesis steps, low sensitivity of antibody analysis, etc., and achieve high sensitivity , The effect of low preparation cost

Inactive Publication Date: 2009-05-20
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In the preparation methods of artificial antigens of pyrethroid pesticides that have been reported abroad, the synthesis conditions of the hapten coupled with the carrier protein are very harsh, and there are many synthesis steps, the acquisition of artificial antigens is difficult and the preparation cost is high; Although the artificial antigens of pyrethroid pesticides are easy to obtain, the analytical sensitivity of the antibodies prepared by them is low

Method used

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  • Pyrethroid pesticide artificial antigen, antibody and preparation thereof
  • Pyrethroid pesticide artificial antigen, antibody and preparation thereof
  • Pyrethroid pesticide artificial antigen, antibody and preparation thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] 1) The hapten synthesis method is as follows:

[0032] Dissolve 1.63g (5mmol) of α-cyano m-phenoxybenzyl alcohol in 5mL of dichloromethane to form a solution with a concentration of 1mol / L, and add it to the reaction flask after being cooled to the reaction temperature; 0.62g (6mmol ) succinic anhydride was dissolved in 2mL of dichloromethane to form a solution with a concentration of 3mol / L, and after being cooled to the reaction temperature, it was added dropwise to the reaction flask. After the dropwise addition, the mixture was stirred and reacted for 12 hours, and the reaction temperature was 14°C. The molar ratio of the reaction raw materials α-cyano-m-phenoxybenzyl alcohol to succinic anhydride is 1:1.2. After the reaction, pyrethroid haptens are generated (where R=CN).

[0033] The structure of the generated type I pyrethroid hapten was identified by electrospray mass spectrometry and nuclear magnetic resonance, and the identification result was: ESI-MS: 324 (...

Embodiment 2

[0046] 1) Synthesize the hapten according to the method in the invention patent "Type I pyrethroid pesticide general hapten and its synthesis method" (application number: ) that the applicant has applied for, and the synthesis method is as follows:

[0047] Dissolve 1.18g (5mmol) of m-phenoxybenzyl alcohol in 5mL of dichloromethane to make a 1mol / L solution, cool to 18°C, and add to the reaction flask after cooling to 18°C; add 1.02g of succinic anhydride ( 10mmol) was dissolved in 10mL of dichloromethane to make a 1mol / L solution, cooled to 18°C ​​and then added dropwise to the reaction flask, and stirred for 15 hours after the dropwise addition, the reaction temperature was 18°C, The molar ratio of benzyl alcohol and succinic anhydride is 1:2. After the reaction, the method of rotary evaporation was used to remove most of the organic solvents, and then the method of silica gel column chromatography was used to purify to obtain 1.47g (4.9mmol) of white solid product, and the ...

Embodiment 3

[0061] 1) Pyrethroid hapten (where R=CN) was synthesized and identified as in step 1) of Example 1.

[0062] 2) Coupling the synthesized hapten compound with protein to prepare artificial antigen, including immune antigen and coating antigen, the method is as follows:

[0063] The hapten synthesized above 0.11g (0.32mmol) (wherein R=CN), equimolar N, N-dicyclohexylcarbodiimide and equimolar N-hydroxysuccinimide were dissolved in 0.6mL of N, N-dimethylformamide, Stir the reaction at 20°C for 4 hours. After the reaction is over, place the reaction mixture in an environment of 4°C overnight (more than 10 hours), then centrifuge, take 0.48mL of the supernatant, divide it into two equal parts, and slowly add it to In 3mL of carbonate buffer solution with a concentration of 15mg / mL hemocyanin, react under stirring for 14 hours. After the reaction is completed, put it into a dialysis bag, and dialyze 8 times with conventional phosphate buffer solution of pH 7.4 as dialysate , wh...

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Abstract

The invention relates to an artificial antigen and an antibody of a pyrethroid pesticide and a preparation method thereof. The artificial antigen is a chemical compound of an artificial antigen of the pyrethroid pesticide, and contains a majority of commonness configuration of the pyrethroid pesticide; the artificial antibody is a polyclonal immunoglobulin G which is prepared by the artificial antigen compound through the immune animal and can react with more than two pyrethroid pesticides for specific immune response. The method has the strengths of easy obtainment of a half antigen of the artificial antigen of the pyrethroid pesticide and environmental friendliness which lower the preparation cost of the artificial antigen; and in the prepared artificial antigen, the common ester bond configuration of the pyrethroid pesticide is retained simultaneously which facilitates the preparation of the high-quality antibody; the antibody has the advantages of high sensitivity and recognizing various pyrethroid pesticides. The antibody can be applied to develop the rapid analytical techniques, methods, and products of multi-residue immunity of the pyrethroid pesticide in the agricultural products and foods.

Description

technical field [0001] The invention relates to a pyrethroid pesticide artificial antigen, antibody and preparation method prepared by selecting a compound having -COOH and containing most of the common structures of pyrethroid pesticides as a hapten. Background technique [0002] The problem of pesticide residues has become a hot issue of general concern in today's society. Pyrethroid pesticides are one of the three major categories of insecticides, which have the characteristics of broad insecticidal spectrum, moderate toxicity, and accumulation. With the prohibition or restriction of highly toxic organophosphorus and carbamate pesticides, the use of pyrethroid pesticides has become more and more widespread, but the problem of such pesticide residues has also become increasingly apparent. The problem of excessive pyrethroid pesticide residues is the most common problem in the export trade of agricultural products such as tea in my country. Therefore, monitoring pyrethroi...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/795C07K14/77C07K16/44C07K16/06
Inventor 李培武张奇张文谢立华丁小霞姜俊陈小媚
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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