Fipronil artificial antigen, antibody and use thereof

An artificial antigen, fipronil technology, applied in the direction of animal/human protein, material inspection product, albumin peptide, etc., can solve the problems of high toxicity and side effects, slow metabolism, high toxicity, etc., and achieve the effect of good affinity and high detection sensitivity

Inactive Publication Date: 2008-01-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, the pesticide is highly toxic (very toxic) to aquatic crustaceans fish, shrimp, and crabs, and also quite toxic (highly toxic) to bees, and metabolizes slowly in the environment. Two metabolites are toxic It is more than...

Method used

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  • Fipronil artificial antigen, antibody and use thereof
  • Fipronil artificial antigen, antibody and use thereof
  • Fipronil artificial antigen, antibody and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1, the preparation of fipronil artificial antigen and antibody:

[0022] A fipronil artificial antigen, the molecular structural formula is (n=3 at this time):

[0023]

[0024] This fipronil artificial antigen is prepared by coupling fipronil hapten FH1 with protein, and the molecular structure formula of the hapten FH1 is: Wherein the binding ratio of the hapten compound to the protein is 5:1-100:1.

[0025] The above-mentioned fipronil hapten FH1 was obtained according to the invention patent "Fipronil Hapten Compound, Synthesis Method and Its Use" that I applied for at the same time. The specific synthesis method is as follows:

[0026] 1) Add 4.51g of 96.7% fipronil technical material (0.01mol) and 50ml of ethylene glycol to a 250ml three-necked flask, heat slowly and stir to dissolve it, and after it is completely dissolved, slowly add 6.92g of 85% concentrated sulfuric acid dropwise, Continue heating and stirring at 138-142°C for reflux reaction for...

Embodiment 2

[0058] Example 2 Establishment and identification of fipronil ELISA method:

[0059] 1. Establishment of Fipronil ELISA assay method and its working conditions and basic parameters

[0060]The hapten and the antibody are respectively covalently coupled with horseradish peroxidase to prepare the enzyme-labeled hapten and the enzyme-labeled antibody. Coat the polystyrene microwell plate with the described coating antigen or antibody, add the mixed solution of the sample to be tested (or fipronil standard sample) and enzyme marker, fipronil, enzyme marker and The coating antigen or antibody on the surface of the well undergoes a competitive immune binding reaction, washing to remove free substances, adding enzyme substrates and chromogenic reagents, and the intensity of the enzymatic color reaction is related to the enzyme label bound to the coating antigen or antibody. It is directly proportional to the amount and inversely proportional to the content of fipronil in the sample ...

Embodiment 3

[0085] Example 3 Preparation example of fipronil direct competition ELISA kit:

[0086] The principle of determination is: firstly, the specific antibody against fipronil is coated on the solid phase carrier, and then the pesticide to be tested and the enzyme-labeled hapten are added, and the pesticide to be tested and the enzyme-labeled hapten compete to bind to the specificity on the solid phase carrier. Antibody, if the content of the pesticide to be tested is large, the enzyme-labeled hapten bound to the solid-phase antibody is less, on the contrary, the enzyme-labeled hapten bound to the solid-phase antibody is more, after the reaction, the substrate is added for color development to be determined. When the enzyme When the amount of labeled hapten is constant, the more the amount of pesticide to be tested is added, the less the enzyme-labeled hapten combined with the solid-phase antibody, the color reaction will be weakened, and the inhibition rate will be increased. On th...

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PUM

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Abstract

A fluo-insect nitrile artificial antigen, its specific antibody there-from and direct or indirect competitive enzyme immune adsorptive determining reagent kit are disclosed. The molecular structural formula is (I), n=1-5, in the (II), n is semi-antigen, and is covalent coupled with protein in proportion of 5:1-100:1 mol, protein is bovine serum albumin or egg-white protein. It can be used to detect fluo-insect nitrile residues in samples.

Description

technical field [0001] The present invention relates to selecting a compound having -COOH and possibly containing the original structure of fipronil as the artificial antigen, artificial antibody and the application of the antibody prepared by the fipronil 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, quantita...

Claims

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Application Information

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IPC IPC(8): C07K14/76C07D231/44G01N33/53
Inventor 金茂俊朱国念程敬丽桂文君
Owner ZHEJIANG UNIV
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