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Synthetic Method for propoxur artificial antigen

A technology of artificial antigen and synthesis method, applied in the biological field, can solve the problems such as the inability to meet the large-scale and rapid detection of pesticide residues, and achieve the effects of simple and feasible antigen preparation technology, strong antigen practicability, and easy factory-scale production.

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

However, relying on these traditional detection methods has been increasingly unable to meet the needs of large-scale and rapid detection of pesticide residues.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Promox Antigen Immunization of New Zealand White Rabbits

[0018] The prepared antigen H 残 -OVA,H 残 -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 th...

Embodiment 2

[0019] Example 2 Competitiveness of propoxur pesticides in combination with propoxur antibody

[0020] The indirect competitive ELISA method was used to conduct a preliminary exploration of the detection curve. Methods as below:

[0021] 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.

[0022] Washing: wash 3 times with PBS

[0023] Blocking: Dilute OVA to 1% in PBS, 200 μl / well, 37°C for 1 hour.

[0024] Washing: wash 3 times with PBS

[0025]Adding samples: Dilute the pesticide step by step by 10 times (the original drug of propoxur is first dissolved with acetone and then diluted with PBS), and diluted to a certain multiple of the corresponding antiserum solution with PBS containing 0.5% OVA. Seconds, mix well) After reacting at room temperature (36°C) for 0.5 hours, add 100 μl / well (equivalent to 50 μl / well of pesticide and antiseru...

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PUM

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Abstract

The present invention relates to synthetic technology of artificial propoxur antigen, and belongs to the field of organic chemistry and immunological technology. The organic chemical process of synthesizing propoxur semi-antigen and antigen includes: synthesis of o-propoxy benzoyl chloride, synthesis of gamma-amino butyric acid, synthesis of artificial propoxur antigen, 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 propoxur antigen with high potency and high compatibility. The present invention has simple preparation process.

Description

(1) Technical field [0001] The method for synthesizing propoxur artificial antigen of the present invention belongs to the field of biotechnology. It is dedicated to the preparation of antibodies that specifically recognize the chemical pesticide propoxur and the development of an immune rapid test kit for the efficient and rapid detection of propoxur content. (2) Background technology [0002] Propoxur (propoxur) belongs to the carbamate chemical insecticide, which was first successfully developed by the German Bayer company, and was introduced in the "Seventh Five-Year Plan" of the country. The propoxur preparation is mainly used for the control of economic crop pests (such as borers on rice, cotton aphids, cabbage aphids, tea orange gall mites, masson pine caterpillars, etc.) and to eliminate various sanitary pests (such as cockroaches, light-colored Culex mosquito, cockroach, etc.), has the characteristics of broad insecticidal spectrum, long residual effect and high ki...

Claims

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Application Information

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IPC IPC(8): C07K1/02C07K14/00G01N33/53G01N33/531
Inventor 刘凤权张奇
Owner NANJING AGRICULTURAL UNIVERSITY
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