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Rhodamine 123 artificial antigen synthesis, antibody preparation method and application

A technology of artificial antigen and synthesis method, applied in chemical instruments and methods, animal/human protein, serum albumin, etc., to achieve the effects of strong specificity, low cost, and easy industrial scale production

Inactive Publication Date: 2009-03-11
HEBEI UNIVERSITY
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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, and is quick and easy to operate without complex pretreatment of samples. The key to immunoassays is the synthesis of rhodamine B artificial antigens and antibodies. Due to the chemical structure of rhodamine B, it is not easy to synthesize artificial antigens, and there are no reports on the detection of rhodamine B.

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  • Rhodamine 123 artificial antigen synthesis, antibody preparation method and application
  • Rhodamine 123 artificial antigen synthesis, antibody preparation method and application

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Embodiment Construction

[0030] The following will be further described in conjunction with the embodiments of the present invention, but not as a limitation of the present invention.

[0031] The overall technical idea of ​​the present embodiment is as follows:

[0032] 1-1. Using rhodamine 123 instead of rhodamine B to synthesize immune antigen:

[0033] Accurately weigh 20-30mg rhodamine 123 and dissolve it in 400μL methanol; accurately weigh 200-250mg bovine serum albumin and dissolve it in 5mL 0.1mol / L, pH7.4 phosphate buffer; slowly add rhodamine 123 methanol solution dropwise Add 3 mL of 25% (V:V) glutaraldehyde aqueous solution to the bovine serum albumin solution, mix well, let stand at room temperature for 1 hour, and freeze at 4°C overnight; Dialyze in 0.01mol / L phosphate buffer for 5 days, then accurately measure the volume of the protein conjugate solution, measure the concentration and binding ratio, add 1 / 1000 sodium azide, and store at 4°C.

[0034] 1-2. Using rhodamine 123 instead o...

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Abstract

The invention belongs to a method for preparing antigens and antibodies, which is particularly used for the specific recognition of inedible pigment rhodamine B and the development of a speed-measuring immune reagent kit of the rhodamine B. The method comprises the synthesis of an artificial antigen of the rhodamine B and a method for preparing an antibody of the rhodamine B. Rhodamine 123 substitutes the rhodamine B to synthesize an immune antigen and cover the antigen, and injects the immune antigen into the body of a New Zealand white rabbit to prepare the high titer antibody of the rhodamine B, and the antibody has good stability and high titer, and solves technical difficulties for the development of an nzyme-linked immune reagent kit of the rhodamine B. The method has strong practicability, does not need high-value equipment during the preparation process of the antigen and the antibody, has strong maneuverability, and is suitable for mass production.

Description

technical field [0001] The invention belongs to the preparation method and application of antigens and antibodies, and is specially used for the specific recognition of non-edible pigment rhodamine B and the development of a rapid test immunological kit for rhodamine B. Background technique [0002] Rhodamine compounds are basic dyes based on xanthene, and there are many homologues. Rhodamine B (Rhodamine B) is a kind of artificially synthesized chemical dye with bright pink color, which has carcinogenic and mutagenic effects. Rhodamine B, commonly known as flower pink, is a basic fluorescent dye. Rhodamine B is fat-soluble and is often used by criminals as condiments (mainly chili powder and chili oil) and as a coloring agent for sausages. Contaminated condiments can cause residues in food preparations. bring great harm to society. Enzyme-linked immunosorbent technology is a new technology developed in the 1960s, and now it has been widely used in rapid and trace detecti...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K16/44G01N33/53
Inventor 王庭欣李小亭赵志磊吴广臣夏立娅刘峥颢庞艳苹
Owner HEBEI UNIVERSITY
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