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Basic orange artificial antigen synthesis and antibody preparation method

A technology of artificial antigen and synthesis method, applied in chemical instruments and methods, animal/human proteins, serum albumin, etc., to achieve the effect of simple and easy preparation technology, strong practicability and good stability

Inactive Publication Date: 2009-04-01
HEBEI UNIVERSITY
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AI Technical Summary

Problems solved by technology

This method can qualitatively and quantitatively detect trace amounts of residual harmful substances in food, and has low requirements for instruments. The key to realize enzyme-linked immunoassay is to synthesize basic orange artificial antigen and antibody when it is applied to the detection report of non-food coloring

Method used

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  • Basic orange artificial antigen synthesis and antibody preparation method
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Embodiment Construction

[0044] The following will be further described in conjunction with the embodiments of the present invention, but not as a limitation of the present invention.

[0045] The overall technical concept of this embodiment is as follows:

[0046] 1. The basic orange artificial antigen synthesis method comprises the following process steps:

[0047] A. Immune antigen synthesis

[0048] Take 150 mg of bovine serum albumin, 15 mg of basic orange, and 1-2 ml of 50% glutaraldehyde solution, and dissolve them in 4-6 ml of phosphate buffer with a concentration of 0.1 mol / liter and pH=7.4 solution, 0.2-0.3 ml of methanol and 1-2 ml of phosphate buffer solution with a concentration of 0.1 mol / L and pH=7.4; using glutaraldehyde as a coupling agent, mix bovine serum albumin solution and basic orange solution Add glutaraldehyde solution dropwise, place at room temperature for 1 hour, place at 4°C for 15-20 hours, centrifuge at high speed for 12-20 minutes, transfer the supernatant to a dialys...

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Abstract

The invention belongs to a preparation method of an antigen and an antibody, which is specially used for recognizing the specificity of inedible pigment basic orange and developing a rapid test immunoassay kit for the basic orange. The invention includes the synthesis of basic orange artificial antigen and a preparation method for an antibody thereof, which mainly comprises: taking glutaric dialdehyde as a coupling agent to synthesize an immunizing antigen and an envelope antigen respectively with bovine serum albumin and ovalbumin, injecting the immunizing antigen to a New Zealand white rabbit to prepare the basic orange antibody with high titer. The antibody has good stability and high titer, and solves the technical difficulty for developing the basic orange enzyme-linked immunoassay kit. The method has the advantages of strong practicality, no need of expensive equipment in the preparation process of the antigen and the antibody and strong operability, thus being suitable for large-scale production.

Description

technical field [0001] The invention belongs to a preparation method of an antigen and an antibody, and is specially used for the specific identification of non-edible pigment basic orange and the development of a rapid test immune kit for basic orange. Background technique [0002] Basic orange is a kind of azo basic dye, the common name is "Wang Jinhuang", the chemical name is 2,4 diamino azobenzene, and the molecular formula is C 12 h 12 N 4 • HCl. Mainly used for dyeing of textiles, leather products and wood products. According to the chemical health and safety database of the National Institute of Health (NIH), ingestion, inhalation, and skin contact of this substance can cause acute and chronic poisoning injuries. Conclusive animal experiment data show that this substance is a carcinogen. According to the "People's Republic of China Hygienic Standards for the Use of Food Additives" (GB2760) and the relevant provisions of the "People's Republic of China Food Sanitat...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K16/44G01N33/53
Inventor 赵志磊李小亭吴广臣王庭欣庞艳苹刘峥颢夏立娅
Owner HEBEI UNIVERSITY
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