Antibody preparation method capable of detecting multiple organophosphorus pesticide residuals

A technology for the preparation of organophosphorus pesticides and antibodies, which is applied in the fields of peptide preparation methods, organic chemistry, chemical instruments and methods, etc., and can solve practical problems such as strong specificity and limited application of immunoassay methods

Inactive Publication Date: 2009-01-14
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to address the deficiencies in the prior art, to provide a method for preparing antibodies that can detect a variety of organophosphorus pesticide residues, and to solve the problem that the antibodies that exist in the current immunological detection of pesticide residues usually have strong specificity to specific substances, and establish Most of the immunoassay methods are only suitable for the detection and analysis of a single pesticide residue, which limits the practical problems of the application of the immunoassay method

Method used

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  • Antibody preparation method capable of detecting multiple organophosphorus pesticide residuals
  • Antibody preparation method capable of detecting multiple organophosphorus pesticide residuals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Such as figure 1 Shown, present embodiment synthesis comprises three steps:

[0032] In the first step, O, O-diethylphosphorothioate was reacted with 4-aminobutyric acid to synthesize a general hapten containing the common structure of various organophosphorus pesticides - O, O-diethylphosphorothioate O,O-diethyl-N-(3-carboxypropyl)thiophosphoramidate (DENP). details as follows:

[0033] Weigh 1.0 g, O, O-diethyl thiophosphoryl chloride dissolved in 5 mL of methanol, placed in an ice bath.

[0034] Weigh 0.69g of KOH and 0.67g of 4-aminobutyric acid, and dissolve them in 5mL of methanol.

[0035] While stirring, add the mixed solution of GABA and KOH dropwise into the O, O-diethylphosphorothioate solution. React in an ice bath for 2 h, filter the reaction mixture, concentrate under reduced pressure, then use 20 ml of NaHCO solution with a concentration of 50 mg / mL to transfer the concentrated solution to a separatory funnel, extract with 10 mL of n-hexane, discard t...

Embodiment 2

[0051] After the antibody was obtained in the same manner as in Example 1, when establishing the method for analyzing the organophosphorus pesticide, except that PBS with 10% methanol was used when diluting the organophosphorus pesticide, the rest was the same as in Example 1. The inhibitory effects of various organophosphorus pesticides on the acquisition of antibodies are shown in Table 3,

[0052] Table 3. IC of several organophosphorus pesticides against the obtained antibodies 50 and IC 20

[0053] Pesticide name

Embodiment 3

[0055] After the antibody was obtained in the same manner as in Example 1, when establishing the method for analyzing the organophosphorus pesticide, except that PBS with 20% methanol was used when diluting the organophosphorus pesticide, the rest was the same as in Example 1. The inhibitory effects of various organophosphorus pesticides on the acquisition of antibodies are shown in Table 4,

[0056] Table 4. IC of several organophosphorus pesticides against the antibodies obtained 50 and IC 20

[0057] Pesticide name

[0058] In summary, it can be seen that the obtained antibody has a strong affinity with the hapten DENP, and also has cross-reactivity with phorate, demeton and diphorate, these three organophosphorus pesticides; Organophosphorus pesticides such as thion and quinthion also had a certain affinity with the obtained antibodies; the antibodies had poor affinity with dimethoate, diazinon and chlorpyrifos.

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Abstract

The invention discloses an antibody preparation method through which various organic phosphorus pesticide residues can be detected and the application, which belongs to the technical filed of agriculture. The antibody preparation method adopts the steps that firstly, O,O-diethylthiophosphoryl chloride is used to react with 4-aminobutyric acid to synthesize into general hapten O, O-diethyl-N-(3-carboxy-propyl)-by-sulfur Phosphoramidate DENP which comprises various organic phosphorus pesticide common structures-O, O-diethyl thiosulfate phosphoryl; secondly, the general hapten O, O-diethyl-N-(3-carboxy-propyl)-by-sulfur Phosphoramidate DENP is respectively coupled with bovine serum albumin BSA and chicken ovalbumin OVA by adopting carbodiimide method, to prepare general immunogen DENP-BSA and general coatingen DENP-OVA; and thirdly, multi-clonal antibody is prepared through immunized New Zealand white rabbits, the obtained antibody has obvious specificity to various organic phosphorus pesticides, such as thimet, demeton, disulfoion, omethoate, phoxim and quinalphos and can be used for the immune detection of the organophosphorus pesticides.

Description

technical field [0001] The invention relates to a preparation method of pesticide residue antibodies in the field of agricultural technology, in particular to a preparation method of antibodies capable of detecting various organophosphorus pesticide residues. Background technique [0002] Pesticides are indispensable substances for the prevention and control of diseases, insects, and weeds in current agricultural production. They play an extremely important role in promoting agricultural production, but at the same time they bring serious environmental problems: ecological imbalance, decline in the quality of agricultural products, and human health. Health is at risk. At present, organophosphorus and pyrethroid pesticides have highly effective insecticidal effects and are the main force of phytochemical protection. With the growing call for environmental protection in recent years, the environmental problems caused by the use of organophosphorus pesticides have attracted in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C07K1/00G01N33/53
Inventor 张敏陆贻通唐建设程国华
Owner SHANGHAI JIAO TONG UNIV
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