Method for synthesis of chlorothalonil artificial antigen

A technology of artificial antigen and synthesis method, which is applied in the direction of chemical instruments and methods, hybrid peptides, ovalbumin, etc., can solve the problems of cumbersome operation, inability to realize rapid detection of a large number of samples, long drug effect period, etc., and achieve simple synthesis steps Effect

Active Publication Date: 2014-06-04
JIANGNAN UNIV
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Problems solved by technology

Chlorothalonil has no systemic conduction effect, but once it is sprayed on the plant body, it can have good adhesion on the surface of the plant and is not easily washed away by rain, so the drug has a long period of efficacy. In addition, it is widely used , therefore, chlorothalonil and its metabolites have relatively large residues in plants and soil, and are even detected in arctic regions
At present, the methods

Method used

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  • Method for synthesis of chlorothalonil artificial antigen
  • Method for synthesis of chlorothalonil artificial antigen
  • Method for synthesis of chlorothalonil artificial antigen

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Embodiment 1

[0032] (1) Synthesis of hapten: Weigh 500mg (1.9mmol) of chlorothalonil, 106.6mg (1.9mmol) of KOH, and 262mg (2mmol) of 6-aminocaproic acid, dissolve these three drugs in 20mL of ethanol, 60℃ React for 12h, then react at 90°C for 12h. Rotate the reactant, then dissolve it with 20mL of water, adjust the pH to 4-5 with hydrochloric acid, produce a precipitate, and dry the precipitate to obtain the hapten CTNH. Identification and analysis were carried out by LC / MS.

[0033] (2) Preparation of conjugate CTNH-BSA

[0034] Weigh 35.8 mg of CTNH, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) 56.9 mg, N-hydroxysuccinimide (NHS) 35.6 mg, and use 1 mL Anhydrous dimethylformamide (DMF) was dissolved (referred to as solution A), and stirred at room temperature for 8 hours. Weigh 112 mg of bovine serum albumin (BSA) and dissolve it in 20 mL of 0.01 mol / L PBS with pH 7.2 (referred to as solution B). Add solution A to solution B drop by drop at room temperature, and react overnight...

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Abstract

The invention discloses a method for synthesis of a chlorothalonil artificial antigen and belongs to the technical field of biochemical engineering. A chlorothalonil raw drug and 6-aminocaproic acid undergo a reaction to produce hapten CTNH as a hydroxy product, the CTNH is coupled with carrier protein bovine serum albumin (BSA) by a carbodiimide method so that a conjugate CTNH-BSA is obtained and is used as an immunizing antigen, the CTNH is coupled with carrier protein chicken egg albumin (OVA) so that a conjugate CTNH-OVA is obtained and is used as a coating antigen, and conjugate coupling ratios are determined by an ultraviolet method. The method realizes successfully synthesis of the chlorothalonil artificial antigen, has simple and effective synthesis steps, can be completely used for immunization analysis and provides a necessary artificial antigen for later research.

Description

technical field [0001] The invention discloses a method for synthesizing an artificial antigen of chlorothalonil, which belongs to the technical field of biochemical industry. Background technique [0002] Chlorothalonil is a broad-spectrum organochlorine fungicide, which can prevent fungal diseases of various crops and green lawns. Chlorothalonil has no systemic conduction effect, but once it is sprayed on the plant body, it can have good adhesion on the surface of the plant and is not easily washed away by rain, so the drug has a long period of efficacy, and it is widely used , therefore, chlorothalonil and its metabolites have relatively large residues in plants and soil, and have even been detected in arctic regions. At present, the methods for detecting chlorothalonil are mainly gas chromatography (GC) and liquid chromatography (HPLC) and other instrumental methods, but these methods have disadvantages such as cumbersome operation, time-consuming, and expensive costs, ...

Claims

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Application Information

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IPC IPC(8): C07K14/77C07K14/765G01N33/68G01N33/53
CPCC07K14/765C07K14/77C07K19/00G01N33/68G01N2430/00
Inventor 匡华胥传来严会娟刘丽强徐丽广马伟宋珊珊
Owner JIANGNAN UNIV
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