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Synthetic method of 4-methylimidazole complete antigen

A technology of methimidazole and complete antigen, which is applied in the direction of animal/human protein, albumin peptide, serum albumin, etc., can solve the problems of high requirements for tested materials, low accuracy, and high cost, and achieve the goal of overcoming direct coupling Slow speed, improved coupling speed and efficiency, and good stability

Active Publication Date: 2012-09-19
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although chromatography and spectrophotometry are simple and quick to operate, they cannot be qualitatively and quantitatively accurate, and the accuracy is low; chromatography can be very accurate for qualitative and quantitative 4-methylimidazole, but it is time-consuming and costly. The requirements for inspection materials are relatively high, and professional and technical personnel are required to operate, which is time-consuming and laborious. It can no longer meet the requirements of modern inspection for convenience, speed and accuracy.
At present, there is no report on the detection of 4-methylimidazole content in food and drug by immunological rapid detection method in China. , on this basis, a complete antigen of 4-methylimidazole that can be used for immunoassay was further designed

Method used

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  • Synthetic method of 4-methylimidazole complete antigen
  • Synthetic method of 4-methylimidazole complete antigen
  • Synthetic method of 4-methylimidazole complete antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Weigh 50 mg of 4-methylimidazole and dissolve it in 5 mL of ethanol, add 250 mg of p-aldehyde benzoic acid to prepare a mixed solution with a mass concentration of 4-methylimidazole of 0.01 g / mL, react at 10°C for 24 hours, and prepare 4 - methylimidazole hapten;

[0028] Dissolve 10 mg of 4-methylimidazole hapten in 10 mL of DMSO to obtain a 4-methylimidazole hapten solution with a concentration of 0.001 g / mL, add 20 mg of EDC and 4 mg of N HS under stirring conditions, and activate for 0.5 h;

[0029] Dissolve 1 mg of protein (BSA or OVA) in 1 mL of phosphate buffer with a concentration of 0.2 mol / L and a pH of 7.0 (0.2 mol / L sodium dihydrogen phosphate and 0.2 mol / L disodium hydrogen phosphate aqueous solution by volume ratio 61:39), prepare a protein solution with a protein concentration of 0.001g / mL, and store it in a refrigerator at 4°C;

[0030] Under the stirring of a magnetic stirrer, add the protein solution dropwise to the activated 4-methylimidazole hapten ...

Embodiment 2

[0045] Weigh 400mg of 4-methylimidazole and dissolve it in 5mL of methanol, add 2g of glyoxylic acid to prepare a mixed solution with a mass concentration of 4-methylimidazole of 0.08g / mL, and react at 30°C for 12h to prepare 4-methylimidazole imidazole hapten;

[0046] Dissolve 50mg of 4-methylimidazole hapten in 5ml DMF to obtain a 4-methylimidazole hapten solution with a concentration of 0.01g / mL, add 50mgEDC and 50mgNHS under stirring conditions, and activate for 6h;

[0047] Dissolve 30 mg of protein (BSA or OVA) in 3 mL of phosphate buffer with a concentration of 0.2 mol / L and a pH of 6.0 (0.2 mol / L sodium dihydrogen phosphate and 0.2 mol / L disodium hydrogen phosphate aqueous solution by volume ratio 12.3:87.7), prepare a protein solution with a protein concentration of 0.01g / ml, and store it in a refrigerator at 4°C;

[0048] Under the stirring of a magnetic stirrer, add the protein solution dropwise to the activated 4-methylimidazole hapten solution to fully react the...

Embodiment 3

[0053] Weigh 2.0 g of 4-methylimidazole and dissolve it in 5 mL of DMSO, add 10 g of p-aldehyde benzoic acid to prepare a mixed solution with a mass concentration of 4-methylimidazole of 0.4 g / mL, and react at 60 ° C for 1 h to prepare 4 - methylimidazole hapten;

[0054] Dissolve 0.5 g of 4-methylimidazole hapten in 1 ml of DMSO to obtain a 4-methylimidazole hapten solution with a concentration of 0.5 g / mL, add 0.4 g of EDC and 2 g of NHS under stirring conditions, and activate for 12 hours;

[0055] Dissolve 5 g of protein (BSA or OVA) in 10 mL of phosphate buffer with a concentration of 0.5 mol / L and a pH of 8.0 (0.5 mol / L sodium dihydrogen phosphate and 0.5 mol / L disodium hydrogen phosphate aqueous solution by volume ratio 94.7:5.3), prepare a protein solution with a protein concentration of 0.5 g / ml, and store it in a refrigerator at 4°C;

[0056] Under the stirring of a magnetic stirrer, add the protein solution dropwise to the activated 4-methylimidazole hapten solutio...

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Abstract

The invention discloses a synthetic method of a 4-methylimidazole complete antigen. The method comprises the following steps of: mixing 4-methylimidazole with an organic acid, dissolving into a solvent, and reacting to obtain a 4-methylimidazole semi-antigen, wherein the organic acid is glyoxalic acid, malonaldehydic acid, succinic semialdehydic acid, pivalic acid, hexuronic acid or p-eormylbenzoic acid; the mass concentration of 4-methylimidazole is 0.01-0.5g / mL; and the mass concentration of the organic acid is 0.05-5g / mL; and combining the 4-methylimidazole semi-antigen with a protein under the activating actions of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and N-hydroxysuccinimide to prepare the 4-methylimidazole complete antigen. According to the method, an artificial antigen of 4-methylimidazole is synthesized successfully, and synthesis steps are simple and effective; and the method can be applied to rapid detection of 4-methylimidazole in foods and medicaments, and necessary artificial antigens are provided for subsequent research, development and application of products.

Description

technical field [0001] The invention relates to 4-methylimidazole complete antigen, in particular to a synthesis method of 4-methylimidazole-protein (BSA and OVA), which belongs to the technical field of biochemical industry. Background technique [0002] 4-Methylimidazole (Chinese alias 4-methylimidazole, 4-methylglycin, 4-methyl-1H-imidazole), English name 4-Methylimidazole (abbreviated 4-Mel or 4-MI), is A nitrogen-containing heterocyclic compound with the molecular formula C 4 h 6 N 2 , is white to off-white crystalline powder, soluble in water and ethanol, corrosive. It is an important pharmaceutical intermediate for the synthesis of various drugs, and it is also an intermediate for the production of various products such as agricultural chemicals, epoxy resin curing agent, dyestuff, industrial emulsifier, metal preservative, etc. 4-Methylimidazole is a by-product in the production of ammonia (ammonium) caramel pigments, which are widely used in wine, beverages, soy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/77
Inventor 于淑娟管永光邓丽卿吴鑫兰
Owner SOUTH CHINA UNIV OF TECH
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