Synthetic method for metolcarb artificial antigen

A technology of antigen synthesis and artificial antigen, which is applied in the biological field to achieve the effects of low cost, simple and feasible antigen preparation technology, and strong antigen practicability

Inactive Publication Date: 2005-05-18
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation of Methiocarb antigen and antibody has not been reported at home and abroad. The structural transformation of hapten and the preparation of antigen are the difficulties and key points of the rapid immunoassay research technology.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] New Zealand White Rabbit Immunized with Methiocarb Antigen

[0020] The prepared antigen H 速 -OVA,H 速 -BSA was used to immunize New Zealand white rabbits over 2kg respectively. The average dosage of each antigen is about 3 mg (based on the amount of protein). For the first immunization, the immunogen emulsified with an equal volume of Freund's complete adjuvant was used, and the rabbit was injected subcutaneously at multiple points; after the first immunization, the immunogen was emulsified with an equal volume of Freund's incomplete adjuvant every 2 weeks. Originally, it was injected subcutaneously or intradermally at multiple points on the back of rabbits and intramuscularly in the hind legs. Beginning from the fourth time, one week after each immunization, blood was collected from the vein of the rabbit's ear to measure the titer (agarose double diffusion method or indirect competition ELISA method). When the titer reaches a certain level (indicated by the maximu...

Embodiment 2

[0021] Example 2 Methiocarb Pesticide Competitiveness Combined with Methiocarb Antibody

[0022] The indirect competitive ELISA method was used to conduct a preliminary exploration of the detection curve. Methods as below:

[0023] Coating: Dilute the coated antigen to a certain number of times with carbonate buffer, add 100 μl per well to a 96-well enzyme-linked micro-analysis plate, and place it overnight at 5°C.

[0024] Washing: wash 3 times with PBS

[0025] Blocking: Dilute OVA to 1% in PBS, 200 μl / well, 37°C for 1 hour.

[0026] Washing: wash 3 times with PBS

[0027] Adding samples: dilute the pesticides step by step by 10 times (the original drug of Methiocarb is first dissolved with acetone and then diluted with PBS), and diluted to a certain multiple with the corresponding antiserum solution containing 0.5% OVA. 30 seconds, fully mixed) after reacting at room temperature (36°C) for 0.5 hours, add 100 μl / well (equivalent to 50 μl / well of pesticide and antiserum s...

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PUM

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Abstract

The present invention relates to synthetic technology of artificial metolcarb antigen, and belongs to the field of organic chemistry and immunological technology. The organic chemical process of synthesizing metolcarb semi-antigen and antigen includes: synthesis of m-methyl benzoyl chloride, synthesis of beta-(m-methyl benzoyl) amino propanoic acid, synthesis of artificial metolcarb antigen, and packing the white artificial metolcarb antigen emulsion in 1 ml centrifugal tubes and preserving at -20 deg.c in refrigerator. The antigen has the features of high stability, powerful immunogenicity, etc. and may be used in preparing metolcarb antigen with high potency and high compatibility. The present invention has simple preparation process.

Description

(1) Technical field [0001] The method for synthesizing an artificial antigen of Methiocarb according to the invention belongs to the field of biotechnology. It is dedicated to the preparation of antibodies that specifically recognize the chemical pesticide medicarb and the development of an efficient and rapid immunoassay for detecting the content of medicarb. (2) Background technology [0002] Metolcarb (metolcarb) belongs to carbamate chemical insecticides and is one of the pesticide varieties produced in large tonnage in my country. Methiocarb preparations have moderate toxicity and are mainly used to control pests in cotton, rice, tea trees, citrus and other crops. They have the characteristics of wide insecticidal spectrum, long residual effect and high killing rate. At present, many manufacturers in China have carried out large-scale production of the original drug or its compound preparation and have been well promoted. [0003] Since the 1970s, due to the prohibiti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/02C07K14/00G01N33/53G01N33/531
Inventor 刘凤权张奇
Owner NANJING AGRICULTURAL UNIVERSITY
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