Organic chlorine pesticide benzoepin artificial antigen and antibody, their preparation and use thereof

A technology of artificial antigen and organochlorine, applied in the field of application in the establishment of immunoassay methods

Inactive Publication Date: 2005-11-23
黄永
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The artificial antigen and specific antibody of endosulfan and the establishment of immunoassay method based on it have not been reported in China

Method used

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  • Organic chlorine pesticide benzoepin artificial antigen and antibody, their preparation and use thereof
  • Organic chlorine pesticide benzoepin artificial antigen and antibody, their preparation and use thereof
  • Organic chlorine pesticide benzoepin artificial antigen and antibody, their preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1. Hapten Synthesis

[0069] Synthesis of Haptens A and B:

[0070] Add succinic anhydride (1-1.5mmol) to 1-hydroxychlordene (1-1.5mmol) in 2 mL of anhydrous pyridine solution, then add dimethylaminopyridine (DMAP), and stir the mixed system overnight. 30 mL of ethyl acetate was added, and the organic layer was washed with acid (HCl), water and brine, and dried over magnesium sulfate. The solution was concentrated, and the residue was passed through a silica gel column (methanol / chloroform / acetic acid 1-3:90-100:0.1-0.5) to obtain product 1 as a white solid.

[0071] 1) Add N-hydroxysuccinimide (0.5-1 mmol) to a solution of compound 1 (0.5-1 mmol) in 10 mL of DCM, and then add DMAP. The mixed system was stirred overnight, filtered and evaporated, and the residue was passed through a silica gel column (acetone / chloroform=1-5:5-10) to obtain a white solid: recrystallized from dichloromethane / petroleum ether to obtain hapten A.

[0072] 2) Treating 1-hydroxychlordene ac...

Embodiment 2

[0110] During the immunoassay, extract the samples using the methanol extraction method ① that is: use 10g of green grapes, carrots, spinach, tea, tobacco and other five samples as the test items, soak in 50ml of methanol and shake for 60 minutes, let it stand, and take it out. solution to obtain the sample to be tested. The resulting soaking solution is the sample to be tested. Others are with embodiment 1.

Embodiment 3

[0112] The detection method adopts the laboratory detection method (the test uses 8-well enzyme labeling strip):

[0113] Coating: Dissolve the prepared antibody in 50 mmol, pH 9.6 carbonic acid buffer to prepare a 10 mg / mL coating solution. Add 100 μl of coating solution to each well of the enzyme-labeled strip overnight. Each well of the coated enzyme-labeled strip was washed three times with PBST (phosphate buffer saline 0.05% (v / v), Tween 20) washing solution.

[0114] Blocking: add 150 μl of 1% bovine serum albumin (BSA) / PBS blocking solution to each well, and block for one hour.

[0115] ·Sampling: Dissolve the sample to be tested in PBS containing 1% BSA, and the endosulfan standard sample is also dissolved in PBS containing 1% BSA. When adding samples, add 50 μl of the above-mentioned sample or standard sample to each well.

[0116] Competitive reaction: dissolve the enzyme-labeled antigen in 0.1% fish skin glue (FG)-PBS buffer, add the sample or standard sample to ...

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Abstract

The invention relates to a pesticides benzoepin artificial antigen-antibody and it's preparing method and application, which specifically relates to preparing small molecular compound artificial hapten, antigen and antibody of cyclopentacliene pesticides benzoepin and application of establishing immunity analysis method. It uses benzoepin derivation as hapten to connect separately with carrier protein like hemocyanin and horse radish peroxidase so as to synthesis artificial antigen and enzyme antigen. Artificial antigen becomes antibody though animal immunity, taking-up blood, antiserum and purity.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry and residue analysis of pesticide small molecule compounds (molecular weight less than 1000 Dalton). In particular, it involves the design and synthesis of artificial haptens and specific antigens of small molecule pesticides such as cyclopentadiene insecticide endosulfan, and the preparation of specific antibodies for immunized animals and their application in establishing immune analysis methods. Background technique [0002] Endosulfan is an extremely broad-spectrum insecticide, mainly targeting a variety of chewing and sucking mouthparts pests, and can also control some moths. Can control Lepidoptera moths and butterfly larvae (such as various leaf-eating, bell-boring, fruit-boring and stem-boring larvae, etc.); Coleoptera beetles and weevils; Homoptera aphids, leafhoppers and whitefly; Stinkbugs of Hemiptera; various thrips of Thysanoptera; midges of Diptera; It is not easy to devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535
Inventor 王硕
Owner 黄永
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