Specific antibody against pesticide meta-tolyl-N-methylcarbamate

A kind of Methiocarb, specific technology, applied in the direction of specific peptides, from serum immunoglobulins, decapeptides, etc., can solve the problems of long detection time, expensive instruments, complicated pretreatment, etc., and achieve good specificity and sensitivity, similar to high degree of effect

A kind of Methiocarb, specific technology, applied in the direction of specific peptides, from serum immunoglobulins, decapeptides, etc., can solve the problems of long detection time, expensive instruments, complicated pretreatment, etc., and achieve good specificity and sensitivity, similar to high degree of effect

CN101747429AInactive Publication Date: 2010-06-23TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Specific antibody against pesticide meta-tolyl-N-methylcarbamate
  • Specific antibody against pesticide meta-tolyl-N-methylcarbamate
  • Specific antibody against pesticide meta-tolyl-N-methylcarbamate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 Meprocarb artificial antigen and antibody preparation

[0042] 1 Pesticide Methiocarb Hapten Synthesis

[0043] (1) Synthesis of hapten 1-(3-carboxyethyl)-3-m-tolylurea (SN)

[0044]

[0045] Weigh 445mg of β-alanine and 665mg of 1-m-toluene isocyanate into a 100mL round bottom flask, add 50mL of anhydrous tetrahydrofuran solution, stir at room temperature overnight, filter, and pass the obtained filtrate through a silica gel column, using a ratio of 1:40 Methanol: dichloromethane was eluted for 1 hour, then the eluent was replaced with pure methanol, the column was washed for 0.5 hours, all the eluate was collected, and vacuum-dried to obtain the Methiocarb hapten 1-(3-carboxyethyl) -3-m-tolylurea.

[0046] (2) Synthesis of 1-(3-carboxyethyl)-3-m-tolylurea activated ester

[0047]

[0048] Take 110mg of the hapten 1-(3-carboxyethyl)-3-m-tolylurea synthesized by the above method, and place 71mg of N-hydroxysuccinimide in a 25ml round bottom flask a...

Embodiment 2

[0060] Embodiment 2 Meprocarb artificial antigen and antibody preparation

[0061] 1 Technical route for the synthesis of the pesticide Methiocarb hapten:

[0062] (1) Synthesis of hapten 1-(3-carboxyethyl)-3-m-tolylurea (SN)

[0063]

[0064] Weigh 460mg of β-alanine and 675mg of 1-m-toluene isocyanate into a 100mL round bottom flask, add 50mL of anhydrous tetrahydrofuran solution, stir at room temperature overnight, filter, and the resulting filtrate is purified by silica gel column chromatography with a ratio of 1: 40 methanol:dichloromethane was eluted for 1 hour, then the eluent was replaced with pure methanol, the column was washed for 0.5 hours, all the eluate was collected, and vacuum-dried to obtain the amethancarb hapten 1-(3-carboxyethyl base)-3-m-tolylurea.

[0065] (2) Synthesis of 1-(3-carboxyethyl)-3-m-tolylurea activated ester

[0066]

[0067] Take 120mg of the hapten 1-(3-carboxyethyl)-3-m-tolylurea synthesized by the above method, and place 73mg of ...

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Abstract

The invention relates to a specific antibody against a pesticide meta-tolyl-N-methylcarbamate and preparation of an artificial antigen and the antibody of the carbamate pesticide meta-tolyl-N-methylcarbamate, in particular to preparation of a hapten, the artificial antigen and the antibody of the carbamate pesticide meta-tolyl-N-methylcarbamate with the molecular structure of 3-methyl benzene. The invention solves the problem that the traditional physical and chemical analysis methods are complicated, high in cost and slow in analysis, and provides simple, convenient, sensitive and accurate immunoassay. In the method, 1-(3-carboxyethyl)-3-m-tolylurea is used as the hapten and is linked with such carrier proteins as hemocyanin and horseradish peroxidase respectively to synthesize the artificial antigen and an enzyme-labelled antigen. The artificial antigen is characterized by simple synthetic method and better stability. The antibody is prepared through animal immunization of the artificial antigen, blood collection and antiserum separation and purification. The antibody is stable, has good specificity and sensitivity, can be used for quick immunoassay of the carbamate pesticide meta-tolyl-N-methylcarbamate and has good application prospect.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry and pesticide residue analysis of small-molecule pesticide compounds (molecular weight less than 1000 Daltons), involving organic synthesis, immunochemistry, biochemistry and other subject knowledge, in particular carbamate with β-alanine structure Design and synthesis of artificial haptens and specific antigens and preparation of specific antibodies of the pesticide-like Methiocarb. Background technique [0002] Since the 1970s, the use of carbamate pesticides has increased year by year due to the successive bans or restrictions on the use of organochlorine pesticides in different countries and the increasing number of insect species resistant to organophosphorus pesticides. With the improvement of living standards and health awareness, people pay more and more attention to the problem of pesticide residues, and more and more attention has been paid to the determination of carbamate pesti...

Claims

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Application Information

Patent Timeline
23 Jun 2010
Publication
CN101747429A
IPC
C07K14/795; C07K16/44; C07K16/06
Inventors
王硕; 孙晶玮