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Synthesis method for atrazine artificial complete antigen

A technology of atrazine and complete antigen, applied in chemical instruments and methods, specific peptides, animal/human proteins, etc., can solve the problems of being expensive and unsuitable for rapid detection, and meet the needs of research, low cost, and synthetic Simple and effective steps

Inactive Publication Date: 2012-03-14
INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

These detection methods require expensive equipment and professional operators, and are not suitable for on-site rapid detection.

Method used

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  • Synthesis method for atrazine artificial complete antigen
  • Synthesis method for atrazine artificial complete antigen
  • Synthesis method for atrazine artificial complete antigen

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Balb / C mouse immunization of embodiment 1 atrazine artificial complete antigen

[0022] Three 6-week-old female Balb / C mice were selected as experimental animals, and AT-BSA was used as the immunogen to immunize the experimental animals. For the first immunization, a single intraperitoneal injection (40 μg·300 μL / mouse) was performed, and the complete antigen was mixed with an equal amount of Freund’s complete For the adjuvant emulsion, the complete antigen and the same amount of Freund's incomplete adjuvant emulsion were used for subsequent immunizations, and the second immunization was performed by intraperitoneal single-point injection (50 μg·300 μL / rat) 14 days later. 14 days later, the third immunization was performed by intraperitoneal single-point injection (70 μg·300 μL / rat), and 20 days later, the fourth immunization was performed by intraperitoneal single-point injection (70 μg·300 μL / rat) with antigen in Freund’s incomplete adjuvant. 3 days before the last fu...

Embodiment 2

[0023] Example 2 The pesticide atrazine competes with the atrazine antibody for binding

[0024] The indirect competitive ELISA method was used as follows:

[0025] Dilute the coating antigen AT-BSA to 2 μg / mL in carbonate buffer, add 100 μL per well to a 96-well enzyme-linked plate, freeze overnight at 4 °C, wash three times with PBST, dilute BSA to 1% in PBS, 120 μL / well, Block at 37°C for one hour, wash with PBST three times, dilute atrazine 2 times and mix it with the corresponding antiserum solution diluted to a certain multiple in equal volume, react at 37°C for 0.5 hour, then add to the blocked enzyme label Add HRP-goat anti-mouse IgG 100μL / well, 1 hour at 37℃, wash 4 times with PBST, develop color with TMB solution for 10 minutes, add 50μL / well 2mol / L sulfuric acid to stop Reaction, reading at the 450nm place of microplate reader, the result shows that antiserum specific competition recognizes atrazine pesticide, can be used for the development of atrazine immune rapi...

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Abstract

The present invention discloses a synthesis method for atrazine (AT) artificial complete antigen, and belongs to the technical field of biochemical engineering. According to the present invention, the structure of the AT is subjected to carboxylation transformation; DCC and NHS are adopted to conjugate the carboxylated AT to a carrier protein BSA; the AT artificial complete antigen is successfully synthesized by ultraviolet spectrum identification and SDS-PAGE identification. With the present invention, the preparation method is simple and effective; the cost is low; the synthesized AT artificial complete antigen can be used for preparing the efficient antibody and developing the rapid immune test kit, and provides a convenient approach for the future detection and research of the AT, wherein the efficient antibody can specially recognize the chemical pesticide AT, the rapid immune test kit can rapidly detect the AT residue.

Description

technical field [0001] The invention relates to a method for synthesizing an artificial complete antigen of atrazine, which belongs to the technical field of biochemical industry, and is used for preparing a high-efficiency antibody that specifically recognizes the chemical pesticide atrazine and developing an immune rapid test that can quickly detect atrazine residues Reagent test kit. Background technique [0002] Herbicide Atrazine (Atrazine, AT) also known as atrazine, full name 2-chloro-4-ethylamine-6-isopropylamine-1,3,5-triazine, molecular weight 215.69, pure product is free Color and odorless crystal, melting point is 175-177°C, solubility in water is 28mg / L at 28°C, solubility in methanol is 1800mg / L at 27°C, solubility in chloroform is 52mg / mL. Since Atrazine was developed and produced by Ciba-Geigy in Switzerland in 1952, it has been widely used in various countries in the world as a selective systemic conduction pre-emergence and post-emergence herbicide, mainly...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K14/795C07D251/52
Inventor 宁保安王彩红柳明王红勇高志贤
Owner INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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