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Chlorogenic acid complete antigen, preparation method and application thereof

A technology of complete antigen and chlorogenic acid, applied in the biological field, can solve the problem of lack of immunogenicity, and achieve the effects of simple and easy preparation method, high specificity and sensitivity, and simple and easy operation of preparation method.

Inactive Publication Date: 2016-01-13
SHENZHEN INST FOR DRUG CONTROL SHENZHEN TESTING CENT OF MEDICAL DEVICES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, chlorogenic acid is a small molecule hapten, which is reactogenic but not immunogenic, and cannot directly stimulate the body to produce an immune response

Method used

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  • Chlorogenic acid complete antigen, preparation method and application thereof
  • Chlorogenic acid complete antigen, preparation method and application thereof
  • Chlorogenic acid complete antigen, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0058] A preparation method of chlorogenic acid complete antigen, comprising the following steps:

[0059] (1) Preparation of the first reaction solution: Weigh 35 mg of chlorogenic acid (CGA), 103 mg of DCC, and 57 mg of NHS and dissolve them in 2 mL of dimethylformamide (DMF), stir and react at room temperature for 4 hours to obtain the first reaction solution;

[0060] (2) Preparation of the second reaction solution: Weigh 20 mg of bovine serum albumin (BSA), dissolve it in 1.2 mL of carbonate buffer at pH 9.6, and place it in an ice bath to prepare the second reaction solution;

[0061] (3) Add 0.6ml of the first reaction solution dropwise to the second reaction solution in an ice bath, stir gently at room temperature, and react for 4 hours to obtain a complete chlorogenic acid antigen mixture.

[0062] Dialysis bag treatment: cut a dialysis bag with a length of 10 cm, boil it with 500 ml of 2% sodium bicarbonate and 1 mmol / L disodium EDTA solution for 10 minutes, wash it ...

Embodiment 2

[0091] A preparation method of chlorogenic acid complete antigen, comprising the following steps:

[0092] (1) Preparation of the first reaction solution: Weigh 35 mg of CGA, 100 mg of DCC, and 50 mg of NHS and dissolve them in 2 mL of dimethylformamide (DMF), stir and react at room temperature for 4 hours to prepare the first reaction solution;

[0093] (2) Preparation of the second reaction solution: 7.5 mg of ovalbumin (OVA), dissolved in 1.2 mL of carbonate buffer solution of pH 9.6, and ice-bathed to obtain the second reaction solution;

[0094] (3) Add 0.6ml of the first reaction solution dropwise to the second reaction solution in an ice bath, stir gently at room temperature, and react for 4 hours to obtain the complete antigen mixture of chlorogenic acid.

[0095] Dialysis bag treatment: cut a dialysis bag with a length of 10 cm, boil it with 500 ml of 2% sodium bicarbonate and 1 mmol / L disodium EDTA solution for 10 minutes, wash it with deionized water, and wash it wi...

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Abstract

The invention provides a chlorogenic acid complete antigen, which is a compound formed by connection of chlorogenic acid to carrier protein through a chemical bond. The invention also provides a preparation method of the chlorogenic acid complete antigen. The method comprises the steps of: (1) dissolving chlorogenic acid and dicyclohexylcarbodiimide, N-hydroxysuccinimide in a solvent, carrying out stirring reaction at room temperature for 2-8h to obtain a first reaction solution, with the molar ratio of chlorogenic acid, dicyclohexylcarbodiimide and N-hydroxysuccinimide being 1:2-5:2-5; (2) dissolving the carrier protein in a buffer solution to obtain a second reaction solution; and (3) adding the first reaction solution into the second reaction solution, performing gentle stirring at room temperature, and carrying out reaction for 2-8h to obtain a mixed solution containing the chlorogenic acid complete antigen, and performing dialysis, thus obtaining the chlorogenic acid complete antigen. The preparation method provided by the invention is simple and easy to operate, and is easy to realize industrial production. The invention also provides application of the chlorogenic acid complete antigen in preparation of drugs or kits for detection of chlorogenic acid.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a complete antigen of chlorogenic acid and its preparation method and application. Background technique [0002] Chlorogenic acid (Chlorogenic acid, CGA) is an important small molecule active ingredient of traditional Chinese medicine, which widely exists in Eucommiaceae, Lonicera genus and Asteraceae plants. Chlorogenic acid is high in Chinese medicinal materials such as capillary, chrysanthemum, honeysuckle, honeysuckle, houttuynia, gardenia, dandelion and thistle, and has antibacterial, antiviral, antioxidative, scavenging free radicals, regulating blood sugar and excitement A variety of biological activities such as the central nervous system have long attracted the attention of the scientific community. Therefore, finding a suitable method for the determination of chlorogenic acid content has become an important link in the quality research of this kind of Chinese medicinal mate...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K14/795C07K14/75C07K14/47C07K14/465C07K1/107G01N33/543
CPCC07K1/1077C07K14/465C07K14/47C07K14/75C07K14/765C07K14/77C07K14/795C07K19/00G01N33/543
Inventor 秦美蓉王平冼静雯王晓炜李俊鹏吴熙
Owner SHENZHEN INST FOR DRUG CONTROL SHENZHEN TESTING CENT OF MEDICAL DEVICES
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