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Method for synthetizing basic orange II artificial antigen

A technology of artificial antigen and synthesis method, applied in animal/human protein, color/spectral property measurement, albumin peptide, etc., to meet the needs of research and the effect of simple synthesis steps

Inactive Publication Date: 2011-07-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing instrumental detection method is difficult to realize the immunodetection of basic orange Ⅱ, and there is no report on the relevant immunological detection method

Method used

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  • Method for synthetizing basic orange II artificial antigen
  • Method for synthetizing basic orange II artificial antigen
  • Method for synthetizing basic orange II artificial antigen

Examples

Experimental program
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Embodiment Construction

[0024] (1) Preparation of artificial hapten

[0025] Solution A: Dissolve 50mg of p-aminobenzenebutyric acid in 2mL of absolute ethanol, adjust the pH to 1~2, add sodium nitrite until the starch potassium iodide test paper turns blue, and react for 1 hour.

[0026] Liquid B: Dissolve 35 mg of m-phenylenediamine in 2 mL of absolute ethanol to prepare Liquid B.

[0027] Add solution A to solution B dropwise, keep the pH at 8-9 during the reaction, and react for 3 hours to prepare the artificial hapten.

[0028] (2) Preparation of complete antigen

[0029] Solution C: Dissolve 8 mg of artificial hapten, 6 mg of EDC, and 4 mg of NHS in 2 mL of PBS (0.01M), and stir magnetically for 3 h.

[0030] Solution D: Dissolve 20mg BSA in 0.05M pH 9.6 carbonate buffer, store at 4°C until use.

[0031] Add solution C dropwise to solution D in an ice bath, and then incubate on ice at 4°C for 3 hours to obtain the artificial antigen mixture.

[0032] Dialysis bag pre-treatment: Take a 10cm ...

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Abstract

The invention relates to a method for synthetizing basic orange II artificial antigen and belongs to the technical field of biochemical industry. The method comprises the following steps of: preparing basic orange II-butyric acid artificial hapten from para-aminophenyl butyric acid and m-phenylenediamine serving as raw materials by a diazotization method; coupling the basic orange II-butyric acidartificial hapten with vector protein bovine serum albumin (BSA) by a carbodiimide method to obtain basic orange II-butyric acid-BSA conjugate; and measuring the coupling ratio of the conjugate by a spectrophotometry. A test result shows that the prepared basic orange II-butyric acid-BSA conjugate generates a good antibody for basic orange II, namely the basic orange II artificial antigen. The basic orange II artificial antigen is synthetized successfully by the method; the synthetic steps are simple and effective; the method can be completely applied to immunoassay; the necessary basic orange II artificial antigen is provided for people to do subsequent researches; and domestic requirements on the researches can be met.

Description

technical field [0001] The invention relates to a synthesis method of basic orange II artificial antigen, which belongs to the technical field of biochemical industry. Background technique [0002] Basic Orange Ⅱ, English name: Basic Orange Ⅱ, CAS: 532-82-1, chemical name 4-phenylazo-1,3-phenylenediamine hydrochloride. Basic orange Ⅱ is an azo basic dye, which is a carcinogen and is forbidden to be used as a food additive. It is mainly used for dyeing textiles, leather products and wood products. According to the National Institute of Health (NIH) chemical health and safety database, it is indicated that excessive intake, inhalation and skin contact with this substance can cause acute and chronic poisoning. The food category that may be added is bean curd skin. The existing instrumental detection method is difficult to realize the immunodetection of basic orange Ⅱ, and there is no report on the relevant immunological detection method. In order to make up for this gap, an ...

Claims

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Application Information

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IPC IPC(8): C07K14/76G01N21/31
Inventor 匡华胥传来宋珊珊林菲赵书阁刘微波赵媛王利兵
Owner JIANGNAN UNIV
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