Artificial antigen, antibody of fenvalerate and uses thereof

A technology of fenvalerate and artificial antigen, which is applied in the direction of animal/human protein, anti-animal/human immunoglobulin, serum albumin, etc., and can solve problems affecting food safety and environmental threats

Inactive Publication Date: 2006-06-21
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, fenvalerate is the most widely used variety of pyrethroid pesticides in my country, whether it is the amount per unit area or the total amount. The excess of fenvalerate residues in agricultural products directly affects food safety and poses a certain threat to the environment.

Method used

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  • Artificial antigen, antibody of fenvalerate and uses thereof
  • Artificial antigen, antibody of fenvalerate and uses thereof
  • Artificial antigen, antibody of fenvalerate and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A kind of fenvalerate artificial antigen, molecular structural formula is (n=2 at this moment):

[0026]

[0027] This fenvalerate artificial antigen is prepared by coupling fenvalerate hapten (QW-PS) to protein, and the molecular structural formula of the hapten is: Wherein the binding ratio of the hapten compound to the protein is 5:1-100:1.

[0028] The above-mentioned fenvalerate hapten (QW-PS) was obtained according to the invention patent "Fenvalerate Hapten Compound, Synthesis Method and Use thereof" which I applied for at the same time. The specific synthesis method is as follows:

[0029] Synthesis of 2-cyano-3-phenoxybenzyl alcohol: Add 7.20g (0.14mol) sodium cyanide and 20ml water to a 250ml two-necked flask, stir to dissolve, add 40ml toluene, 20.60g (0.1mol) m-benzene Oxybenzaldehyde, 0.65g tetrabutylammonium bromide, add 15ml of 36-38% hydrochloric acid dropwise at room temperature, continue to react for 1.5hr after adding, then add 12.5ml of water, dis...

Embodiment 2

[0062] A kind of fenvalerate artificial antigen, molecular structural formula is (n=5 at this moment):

[0063]

[0064] This fenvalerate artificial antigen is prepared by coupling fenvalerate hapten (QW-He) with protein, and the molecular structural formula of hapten is: Wherein the binding ratio of the hapten compound to the protein is 5:1-100:1.

[0065] The above-mentioned fenvalerate hapten (QW-He) was obtained according to the invention patent "Fenvalerate Hapten Compound, Synthesis Method and Use thereof" which I applied for at the same time. The specific synthesis method is as follows:

[0066] Synthesis of 2-cyano-3-phenoxybenzyl alcohol: Add 7.20g (0.14mol) sodium cyanide and 20ml water to a 250ml two-necked flask, stir to dissolve, add 40ml toluene, 20.60g (0.1mol) m-benzene Oxybenzaldehyde, 0.65g of tetrabutylammonium bromide, cooled to below 0 degree with ice-water bath, dropwise added 15ml of 36-38% hydrochloric acid, controlled the speed, finished dropping ...

Embodiment 3

[0084] Example 3 Establishment and identification of fenvalerate ELISA method

[0085] 1. Establishment of ELISA assay method for fenvalerate and its working conditions and basic parameters

[0086] Direct competition enzyme-linked immunoassay method was used. The principle of its determination is that the compound obtained by coupling the pesticide molecule with the macromolecule carrier (such as protein) is adsorbed on the solid phase carrier (96-well microtiter plate) as the coating antigen to prepare the solid phase antigen, and then add the compound to be tested. Pesticides and corresponding enzyme-labeled antibodies. The solid-phase antigen, the pesticide to be tested, and the enzyme-labeled antibody undergo a competitive binding reaction. If the content of the pesticide to be tested is large, the enzyme-labeled antibody bound to the solid-phase antigen is less, and on the contrary, there are more enzyme-labeled antibodies bound to the solid-phase antigen. The substrat...

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Abstract

The invention discloses a fenvalerate artificial antigen, with the molecular structural formula is as formulaó±, n=1-5, employing formula ó� as partial antigen, covalent coupling with protein for synthesis; the combination ratio between the partial antige and protein is 5:1-100:1. The invention also discloses the monoclonal or polyclonal immune globulin G which is got from immunizating mouse or rabbit by the above said fenvalerate artificial antigen and can occur specific immunological reaction with fenvalerate, the said fenvalerate specific antibody can be used to check the residual quantity of fenvalerate in sample. The invention also discloses a detecting agent box of direct and indirect competeing enzyme and immunoadsorption for fenvalerate residual analysis. The got antibody is used to detect fenvalerate by ELISA method, with the lowest detecting limit being 4.0í‚1.5 ug / l (0.004ppm), and the detecting sensitivity is high.

Description

technical field [0001] The present invention relates to selecting a compound having -COOH and possibly containing the original structure of fenvalerate as the artificial antigen, artificial antibody and the application of the antibody prepared by fenvalerate hapten. Background technique [0002] The invention belongs to the technical field of immunochemistry and residue analysis of small-molecule pesticide compounds (molecular weight less than 1000 Daltons), relates to organic synthesis, immunochemistry and biochemistry, and relies on basic principles of immunology and immunochemistry and biotechnological means to design and synthesize Small molecule target analyte hapten, and coupled with carrier protein to prepare effective artificial antigen, immunize animals to prepare specific antibody for small molecule analyte, use the specific immunological reaction of antigen and antibody and the identification of markers that are easily detected and identified Amplification, quanti...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07C235/34C07K14/77C07K16/18G01N33/53
Inventor 朱国念金仁耀程敬丽桂文君陈则利金茂俊王春梅
Owner ZHEJIANG UNIV
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