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Preparation of triazoline artificial semi-antigen, antigen and antibody

A technology of artificial hapten and triazophos, which is used in the preparation of antibodies and the field of antigens, can solve the problems of cumbersome, complicated and toxic processes, and achieve the effects of high sensitivity, good affinity and low cross-reaction rate.

Inactive Publication Date: 2005-10-12
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Triazophos original drug is moderately toxic and toxic to fish and bees
However, this method requires complex instruments that most laboratories do not have, and the process is cumbersome, so it is not suitable for screening and testing of large batches of samples

Method used

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  • Preparation of triazoline artificial semi-antigen, antigen and antibody
  • Preparation of triazoline artificial semi-antigen, antigen and antibody
  • Preparation of triazoline artificial semi-antigen, antigen and antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Triazophos artificial hapten synthesis method is as follows:

[0038] 1) Synthesis of O-ethylthiophosphoryl dichloride (TZLM-1)

[0039] Weigh the phosphorus trichloride (PSCl 3 ) 68g (about 0.4mol) was placed in a three-necked flask with a low-temperature thermometer, cooled to -10~-5°C with an ice-salt water bath, and 18.4g (about 0.4mol) of absolute ethanol was added dropwise under vigorous stirring, Strictly control the rate of addition so that the temperature of the reaction solution is always at -30 to 0°C. After the dropwise addition was completed, the reaction was continued at -20-10°C for 1 h. After the reaction, the reaction solution (100ml×2) was washed with (0±5)°C distilled water, the oil layer was separated and washed with anhydrous Na 2 SO 4 Dry it, then distill it under reduced pressure with a water pump, and collect fractions at 65-75° C. to obtain 28.5 g of a colorless transparent oily liquid (yield 40%, calculated as phosphorus trichloride).

...

Embodiment 2

[0074] 1. Triazophos artificial hapten synthesis method is as follows:

[0075] 1) Synthesis of O-ethylthiophosphoryl dichloride (TZLM-1)

[0076] Weigh the phosphorus trichloride (PSCl 3 ) 68g (about 0.4mol) was placed in a three-necked flask with a low-temperature thermometer, cooled to -10~-5°C with an ice-salt water bath, and 55g (about 1.2mol) of absolute ethanol was added dropwise under vigorous stirring, strictly Control the rate of addition so that the temperature of the reaction solution is always at -30 to 0°C. After the dropwise addition, the reaction was continued at -20~10°C for 2h. After the reaction is complete, wash the reaction solution (100ml×2) with (0±5)°C distilled water, separate the oil layer and wash it with anhydrous Na 2 SO 4 After drying, it was distilled under reduced pressure by a water pump, and the fraction at 65-75°C was collected to obtain 51.8 g of a colorless transparent oily liquid (yield 72.3%, calculated as phosphorus trichloride).

...

Embodiment 3

[0111] 1. The synthetic method of triazophos artificial hapten is as follows:

[0112] 1) Synthesis of O-ethylthiophosphoryl dichloride (TZLM-1)

[0113] Weigh the phosphorus trichloride (PSCl 3 ) 68g (about 0.4mol) was placed in a three-necked flask with a low-temperature thermometer, cooled to -10~-5°C with an ice-salt water bath, and 92g (about 2.0mol) of absolute ethanol was added dropwise under vigorous stirring, strictly Control the rate of addition so that the temperature of the reaction solution is always at -30 to 0°C. After the dropwise addition, the reaction was continued at -20-10°C for 5h. After the reaction, the reaction solution (100ml×2) was washed with (0±5)°C distilled water, the oil layer was separated and washed with anhydrous Na 2 SO 4 After drying, it was distilled under reduced pressure by a water pump, and the fraction at 65-75°C was collected to obtain 57.0 g of a colorless transparent oily liquid (yield 79.53%, calculated as phosphorus trichloride...

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Abstract

Production of triazophosphous artificial semi-antigen, antigen and antibody is carried out by taking O-ethyl thiophsphoryl dichloride as material, reacting O-ethyl thiophsphoryl dichloride with synthetic intermediate 1-phenyl-1,2,4-triazoalcohol of triazophosphous under catalyst action, generating monochloride, reacting monochloride with propalanine under acid-binding agent action, generating triazophosphous semi-antigen TZLBu, reacting monochloride with amino acetic acid under acid-binding agent action, generating TZLHe, coupling with protein by carbodi-imide method and mixed acid anhydride method, and obtaining artificial antigen. Its advantages include rapid, convenient, low cost, and specific antibody with high affinity.

Description

technical field [0001] The present invention relates to a triazophos hapten of a compound having -COOH and possibly containing the original structure of triazophos, as well as a method for preparing the corresponding antigen and antibody. Background technique [0002] The invention belongs to the technical field of immunochemistry and residue analysis of small-molecule pesticide compounds (molecular weight less than 1000 Daltons), relates to organic synthesis, immunochemistry and biochemistry, and relies on basic principles of immunology and immunochemistry and biotechnological means to design and synthesize Small molecule target analyte hapten, and coupled with carrier protein to prepare effective artificial antigen, immunize animals to prepare specific antibody for small molecule analyte, use the specific immunological reaction of antigen and antibody and the identification of markers that are easily detected and identified Amplification, quantitative detection of ultra-tr...

Claims

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Application Information

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IPC IPC(8): C07F9/6518C07K1/107C07K16/18C07K17/02
Inventor 朱国念桂文君程敬丽
Owner ZHEJIANG UNIV
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