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Synthesis method of artificial antigen suitable for aminoazobenzene

A technology of p-aminoazobenzene and aminoazobenzenebutyric acid, which is applied in the field of biochemical industry, can solve the problems of undetectable and few aminoazobenzene enzyme-linked immunoassays, etc., and achieve the effect of simple synthesis steps

Inactive Publication Date: 2012-12-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the limitations of detection technology, conventional inspection methods cannot detect it
In recent years, studies on azo-based immunoassay methods have been carried out at home and abroad, but there are few reports on enzyme-linked immunoassays for p-aminoazobenzene at home and abroad. It is necessary to prepare antibodies that can produce better specific antibodies. Aminoazobenzene artificial antigen

Method used

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  • Synthesis method of artificial antigen suitable for aminoazobenzene
  • Synthesis method of artificial antigen suitable for aminoazobenzene
  • Synthesis method of artificial antigen suitable for aminoazobenzene

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Experimental program
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Embodiment Construction

[0025] (1) Preparation of p-aminoazobenzene derivatives

[0026] The substrate p-aminophenylbutyric acid (358 mg, 2 mmol) was dissolved in dilute hydrochloric acid (1N, 4 mL) under ice bath (-5°C-0°C), and then sodium nitrite (138 mg, 2 mmol) was added , after stirring for 15 minutes, add aniline (186 mg, 2 mmol), keep the whole process below 0°C for 0.5h, the reaction monitoring (LC / MS) shows that the raw materials are completely consumed, and the reaction solution is transferred to preparative chromatography for purification, and p-aminoazo Benzene Derivatives 1.

[0027] Similarly, p-aminoazobenzene derivative 2 can be obtained by replacing the substrate with p-aminophenylacetic acid.

[0028] (2) Preparation of p-aminoazobenzene artificial antigen

[0029] Take 0.5mL of N,N-dimethylformamide DMF / water solution containing 0.1mmol p-aminoazobenzene derivatives, add 0.15mmol carbodiimide EDC and 0.15mmol N-hydroxysuccinimide NHS at room temperature and avoid light Reaction...

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Abstract

The invention relates to a synthesis method of an aminoazobenzene artificial antigen, belonging to the technical field of biochemical engineering. The method comprises the following steps: respectively diazotizing p-aminophenylbutyric acid and p-aminophenylacetic acid and reacting with aniline to obtain two derivatives 1 and 2 for aminoazobenzene; meanwhile, coupling the aminoazobenzene derivative 1, which is used as a hapten, with a carrier protein BSA (bovine serum albumin) by an active ester process to obtain an immunogen for immunizing a rabbit to generate an antibody; coupling the aminoazobenzene derivative 2, which is used as a hapten, with a carrier protein OVA (ovalbumin) by an active ester process to obtain a coating antigen; and determining the coupling ratio of the coupling substance with an ultraviolet scanner. The invention successfully synthesizes the aminoazobenzene artificial antigen, has the advantage of simple and effective synthesis steps, obtains an favorable antibody for aminoazobenzene specificity when being used in immunoassay, and can satisfy the demands for research in China.

Description

technical field [0001] The invention relates to a method for synthesizing a universal artificial antigen applicable to p-aminoazobenzene, which belongs to the technical field of biochemical engineering. Background technique [0002] p-aminoazobenzene, English name: p-aminoazobenzene; Aniline Yellow, 4-Aminoazobenzene; chemical name is also called 4-aminoazobenzene, aniline yellow; CAS number: 60-09-3; molecular formula is C 12 h 11 N 3 . Azo dyes (a class of organic compounds in which the two ends of the azo group are connected to aryl groups) are the most widely used synthetic dyes in the printing and dyeing process of textiles and clothing. Under special conditions, it can decompose and produce more than 20 carcinogenic aromatic amines. Change the DNA structure of the human body through activation to cause pathological changes and induce cancer. Azo is one of the mandatory inspection items required by international environmental protection. There are three inspection m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/77
Inventor 胥传来李灼坤匡华王风艳王利兵刘丽强胡拥明
Owner JIANGNAN UNIV