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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: 2007-08-15
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, the molecular structural formula is (in this case, n=2):

[0026]

[0027] This fenvalerate artificial antigen is prepared by coupling the fenvalerate hapten (QW-PS) and protein. The molecular structure of the hapten is: The binding ratio of hapten compound to 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 its use" that I applied 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 into a 250ml two-necked flask. After stirring to dissolve, add 40ml toluene and 20.60g (0.1mol) m-benzene Oxybenzaldehyde, 0.65g tetrabutylammonium bromide, 15ml of 36~38% hydrochloric acid was added dropwise at room temperature, after the addition, the reaction was continued for 1.5hr, and then 12.5ml of water was...

Embodiment 2

[0063] A kind of fenvalerate artificial antigen, the molecular structural formula is (in this case, n=5):

[0064]

[0065] This fenvalerate artificial antigen is prepared by coupling the fenvalerate hapten (QW-He) and protein. The molecular structure of the hapten is: The binding ratio of hapten compound to protein is 5:1-100:1.

[0066] The above-mentioned fenvalerate hapten (QW-He) was obtained according to the invention patent "Fenvalerate hapten compound, synthesis method and its use" which I applied at the same time. The specific synthesis method is as follows:

[0067] Synthesis of 2-cyano-3-phenoxybenzyl alcohol: Add 7.20g (0.14mol) sodium cyanide and 20ml water into a 250ml two-necked flask. After stirring to dissolve, add 40ml toluene and 20.60g (0.1mol) m-benzene Oxybenzaldehyde, 0.65g tetrabutylammonium bromide, cooled to below 0 degrees with an ice water bath, dropwise add 15ml of 36~38% hydrochloric acid, control the speed, about 30min to drip, continue to react f...

Embodiment 3

[0086] Example 3 Establishment and identification of fenvalerate enzyme-linked immunosorbent assay method

[0087] 1. Establishment of fenvalerate ELISA method and its working conditions and basic parameters

[0088] Using direct competition enzyme-linked immunosorbent assay method. The principle of the determination is that the compound prepared by coupling the pesticide molecule and the macromolecular carrier (such as protein) as the coated antigen is adsorbed on the solid-phase carrier (96-well microtiter plate) to prepare the solid-phase antigen, and then add it to the test 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 high, the enzyme-labeled antibody bound to the solid-phase antigen will be less. On the contrary, there are more enzyme-labeled antibodies bound to the solid-phase antigen. The subst...

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Abstract

The invention discloses a fenvalerate artificial antigen, with the molecular structural formula is as formula I, n=1-5, employing formula II 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 4.0+ / -1.5 ug / l (0.004ppm), and the detecting sensitivity is high.

Description

Technical field [0001] The invention relates to the use of selecting a compound with -COOH and most likely to contain the original structure of fenvalerate as an artificial antigen, artificial antibody and antibody prepared by fenvalerate hapten. Background technique [0002] The invention belongs to the technical field of immunochemistry and residue analysis of pesticide small molecule compounds (molecular weight less than 1000 Daltons), involving organic synthesis, immunochemistry and biochemistry, etc., relying on the basic principles of immunology, immunochemistry and biotechnological means to design and synthesize The small molecule target analyte hapten is coupled with the carrier protein to prepare effective artificial antigens. The immunized animal prepares specific antibodies to the small molecule analyte, using the specific immunological reaction of the antigen antibody and the identification of the markers that can be easily detected. Amplification, quantitative detect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K14/77C07K16/18G01N33/53C07C235/34
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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