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Preparation method for artificial antigen of theophylline

An artificial antigen, artificial hapten technology, applied in the direction of animal/human protein, albumin peptide, serum albumin, etc., can solve the problems of expensive instruments, time-consuming detection, inability to achieve, and achieve advanced synthesis technology and strong specificity. Effect

Active Publication Date: 2013-10-23
杭州博林生物技术有限公司
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Problems solved by technology

[0005] At present, the detection of theophylline mainly relies on high-performance liquid chromatography (HPLC), gas chromatography (GC), thin-layer chromatography (TLC), simple and simple (MS), etc., but the equipment is expensive, the detection is time-consuming, and professional and technical personnel are required. The operation cannot meet the fast and accurate requirements of modern detection

Method used

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  • Preparation method for artificial antigen of theophylline
  • Preparation method for artificial antigen of theophylline
  • Preparation method for artificial antigen of theophylline

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Effect test

Embodiment 1

[0032] (1) Preparation of artificial hapten:

[0033] (a) Add 500mg of 5,6-diamino-1,3-dimethyluracil (1) into a round bottom flask, add 10ml of DMF solution, then add 500mg of glutaric anhydride, and stir the reaction at room temperature for 17 hour; after the reaction finished, the reaction solution was dried with an oil pump to obtain 942mg yellow solid product (2); TLC detection: the chromatographic solution was (absolute ethanol: strong ammonia water: CHCL 2 ︰1,4dioxane=8︰1︰10︰1, v / v) product R f =0.2.

[0034] (b) Add 15ml of 2N NaOH solution to the product of the previous step, and reflux in an oil bath at 75°C for 1.5 hours. After the reaction, cool down and adjust the pH to 3 with 6N hydrochloric acid. A large amount of precipitation occurs, and the solid is recovered by filtration. Then recrystallized by hot water to obtain 240mg solid product (3); TLC detection: the chromatographic solution is (absolute ethanol: concentrated ammonia water: CHCL 2 ︰1,4dioxane=8...

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Abstract

The invention provides a preparation method for an artificial antigen of theophylline. The preparation method comprises the following steps of: (a) preparing and detecting a hapten: reacting 5,6-diamido-1,3-dimethylurea pyrimidine (1) with glutaric anhydride to obtain the hapten containing carboxyl; (b) preparing and detecting the artificial antigen: combining the hapten with bovine serum albumin (BSA) by utilizing a carbodiimide method, so as to prepare the artificial antigen of the theophylline, namely theophylline-bovine serum albumin. According to the preparation method, the prepared artificial antigen of the theophylline can be subjected to animal immunization to obtain corresponding theophylline antibodies and can be used for researching various theophylline immunoassay methods, and a relatively convenient, fast and accurate way is provided for the detection of the theophylline.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a preparation method of theophylline artificial antigen. Background technique [0002] Theophylline (Theophylline) Alias: Dioxodimethylpurine; Theophylline hydrate; Theophylline; Theophylline is a methyl purine drug. It has the effects of strengthening the heart, diuresis, expanding coronary arteries, relaxing bronchial smooth muscle and stimulating the central nervous system. Mainly used for the treatment of bronchial asthma, emphysema, bronchitis, cardiac dyspnea. Its structural formula is: [0003] [0004] Theophylline is a central stimulant, which can enhance the exciting process of the cerebral cortex and also excite the myocardium. Theophylline is mainly used clinically to treat obstructive respiratory disorders and asphyxia of premature infants. Excessive use will lead to excessive myocardial excitement, cardiac convulsions, and a sharp drop...

Claims

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Application Information

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IPC IPC(8): C07K14/765
Inventor 王百龙邵越水
Owner 杭州博林生物技术有限公司
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