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Method for synthesizing rhodamine B complete antigen

A technology of complete antigen and synthesis method, which can be applied in the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problem of difficult rhodamine B residues and other problems, and achieve the effect of concise synthesis steps.

Inactive Publication Date: 2009-11-25
JIANGNAN UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

Existing instrumental detection methods are difficult to achieve rapid, accurate and multi-residue detection of Rhodamine B
At present, there is no ideal report on the multi-residue immunoassay method for rhodamine B at home and abroad

Method used

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  • Method for synthesizing rhodamine B complete antigen

Examples

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Embodiment 1

[0019] (1) Preparation of rhodamine B complete antigen

[0020] Preparation of liquid A: 20 mg of rhodamine B was dissolved in 1 mL of DMF. After complete dissolution, 10 μL of tributylamine was added under ice bath to mix evenly, and then 10 μL of isobutyl chloroformate was stirred for 1 hour by magnetic force.

[0021] Preparation B solution: 40mg BSA was dissolved in 3mL pH 7.4 phosphate buffer, and stored at 4°C for later use.

[0022] Add liquid A dropwise to liquid B under ice-cooling, and then incubate at 4°C for 3 hours. The rhodamine B complete antigen mixture is obtained.

[0023] Dialysis bag pre-treatment: Take a 10cm dialysis bag, boil it in boiling water for 5 minutes, rinse it with 60°C deionized water for 3 minutes, and store it in 4°C deionized water for later use.

[0024] Dialysis: transfer the rhodamine B complete antigen mixture into a dialysis bag, and dialyze with 2×2L of 0.01M PBS solution of pH 7.4 and 2×2L of deionized water for 3 days. Finally, th...

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Abstract

The invention discloses a method for synthesizing rhodamine B complete antigen, and belongs to the technical field of biochemical engineering. The method takes rhodamine B as a hapten, couples the rhodamine B with carrier protein BSA by a mixed acid anhydride method, and measures the coupling ratio of a conjugate by a spectrophotometry. The method successfully synthesizes the complete antigen of rhodamine B, has simple and effective synthesis steps, can be fully used in immunoassay, provides necessary artificial antigen for future research of people, and can meet the requirement of national rhodamine B research.

Description

technical field [0001] The invention discloses a method for synthesizing complete rhodamine B antigen, which belongs to the field of biochemical technology. Background technique [0002] Rhodamine B, English name: Rhodamine B; CAS: 81-88-9, molecular formula C 28 h 31 ClN 2 o 3 Rhodamine B is a kind of artificially synthesized dye with bright pink color, which has strong fluorescence in solution, and is used as cell fluorescence staining agent in the laboratory, colored glass, characteristic fireworks and firecrackers and other industries. It is a triphenylmethane basic dye with potential carcinogenicity and mutagenicity. It is not allowed to be used in food in my country and the European Union. Rhodamine B is fat-soluble and is used as a coloring agent for condiments (mainly chili powder and chili oil). Contaminated condiments can cause residues in foods made with them. The condiment has a high content when dyed with rhodamine B, and it is even directly mixed in, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K1/34C07K1/113
Inventor 胥传来宋珊珊林菲
Owner JIANGNAN UNIV
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