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Monoclonal antibody for O, O-dimethoxythiophosphate pesticides

A technology of dimethoxyphosphorothioate and dimethylphosphorothioate, applied in the fields of genetic engineering, plant gene improvement, biochemical equipment and methods, etc., to achieve easy popularization and promotion, large analysis capacity, and easy operation simple effect

Active Publication Date: 2009-11-04
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The multi-residue ELISA detection method for detecting multiple pesticides at the same time has been reported in foreign countries in the 1990s. Only in my country in recent years have units and researchers paid attention to and engaged in the research of multi-residue pesticide detection technology. At present, there is no such method in China. research report

Method used

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  • Monoclonal antibody for O, O-dimethoxythiophosphate pesticides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Detect O in the tap water sample of embodiment 1, O-dimethoxy phosphorothioate (taking methyl parathion as an example) pesticide residue

[0052] Main conditions of ELISA reaction system

[0053] "3-Methyl 4-nitrophenyl-O-butyric acid-ovalbumin conjugate" coating 2.5μg / ml was added to each well of the microanalysis plate in 50μl, and the monoclonal antibody was diluted 32 times as the working concentration , 1% gelatin as a blocker, the methanol content in the antigen-antibody reaction solution phosphate-Tween buffer is 5%, the pH value is 7.4, the ionic strength is 0.15mol / L, and each hole in the microanalysis plate is 50 μl.

[0054] Sample preparation for testing

[0055] Tap water sample: Measure 1L of tap water, add 2mg of methyl parathion standard standard net weight, and prepare a 2ppm liquid sample.

[0056] The above samples were detected by ELISA detection method, and the operation was as follows:

[0057] (1) Coated

[0058] "3-Methyl 4-nitrophenyl-O-buty...

Embodiment 2

[0078] Detect O in the peach fruit sample of embodiment 2, O-dimethoxy phosphorothioate (taking methyl parathion as an example) residual

[0079] The main conditions of the ELISA reaction system are the same as in Example 1.

[0080] Sample preparation for testing

[0081] Peach fruit sample: Weigh 10 g of chopped peach fruit, add 0.01 mg net weight of methyl parathion, and prepare a solid sample of 1 ppm.

[0082] The above samples were detected by ELISA detection method, and the operation was as follows:

[0083] Steps (1), (2) are the same as in Example 1.

[0084] (3) Add antibody and the mixture of antibody and sample to be tested

[0085] Weigh 0.1 g of the sample to be tested, soak it in 0.2 ml of methanol for about 1 hour, take 100 μL and add it to 400 μL of phosphate buffer containing 0.5% Tween 20 to prepare the test solution, and dilute the cell culture supernatant 16 times with the test solution The test solution and the phosphate buffer containing 0.5% Tween 2...

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Abstract

The invention relates to a monoclonal antibody for O,O-dimethoxythiophosphate pesticides and a preparation method thereof, which belong to the biotechnical field. An immunogen, namely a 3-[4-(O,O-dimethylphosphorothioyloxy)phenyl]-1-propionic acid-bovine serum albumin conjugate is used to immunize a BALB / C mouse, and spleen cells and Sp2 / 0 myeloma cells are used to prepare hybridoma cells through hybridoma technology, thereby obtaining the steady secretion of the monoclonal antibody against the O,O-dimethoxythiophosphate pesticides. The invention relates to preparation of the monoclonal antibody special for identifying the O,O-dimethoxythiophosphate pesticides, and the high-sensitivity quick detection thereof on pesticide residue in agricultural products and agricultural production environment. The preparation technology of the antibody is simple, convenient and feasible: the whole preparation process of an antigen does not need special instruments and equipment, and is easy for mass production of factories.

Description

(1) Technical field [0001] The invention relates to an O, O-dimethoxy phosphorothioate pesticide monoclonal antibody and a preparation method thereof, belonging to the field of biotechnology. Dedicated to the multi-residue detection of pesticides in agricultural products and agricultural production environments, especially suitable for large-scale sample detection and on-site monitoring. (2) Background technology [0002] At present, organophosphorus pesticides are the most widely used pesticides in my country. Organophosphorus pesticides have many control objects and a wide range of applications. They play an important role in preventing and controlling crop diseases, insects, and weeds, and ensuring high crop yields and harvests. Pollution also causes serious harm to human health. In recent years, the Ministry of Agriculture has successively announced a batch of 23 kinds of pesticides banned by the state, including 5 kinds of highly toxic organophosphorus pesticides incl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C12N15/06C12P21/08C12N5/20
Inventor 刘凤权刘艳娄旸
Owner NANJING AGRICULTURAL UNIVERSITY
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