Application of a composition and test kit thereof in detecting glycocholic acid

A kit and a technique for glycocholic acid, which are applied in the field of medical detection, can solve problems such as cumbersome processes, and achieve the effects of avoiding cumbersome processes, simple preparation procedures and high sensitivity.

CN113721012BActive Publication Date: 2022-08-09SHENZHEN AMTECH BIOENGINEERING LTD INC
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2022-08-09

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Abstract

The application of a composition and a kit thereof in detecting glycocholic acid, the composition comprising a glycocholic acid-biotin derivative formed by combining glycocholic acid and biotin. The present invention designs a glycocholic acid detection kit based on an immunoturbidimetric platform, wherein the glycocholic acid-biotin small molecule synthesis method is fast and simple, and the combination of biotin-avidin is used to design a kit. The new reaction principle overcomes the shortcomings of the existing technology. The glycocholic acid-biotin small molecule does not need to be combined with macromolecular proteins or microspheres, and is directly applied to the latex immunoturbidimetric platform as a competitor, avoiding the preparation of macromolecular competitors. tedious process.
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Description

technical field

[0001] The invention relates to the technical field of medical detection, in particular to the use of a composition and a kit thereof in the detection of glycocholic acid. Background technique

[0002] Cholyglycine (CG) is a conjugated cholic acid formed by combining cholic acid and glycine. Glycolic acid is mainly synthesized in the liver and is excreted into the intestine with bile. Under normal circumstances, the concentration of Glycolic acid in the blood is extremely low.

[0003] The structural formula of Glycolic acid is as follows:

[0004]

[0005] When liver cells are damaged, the ability of hepatocytes to absorb glycocholic acid decreases, resulting in an increase in the content of glycocholic acid in the blood; when bile is stagnant, the liver excretion of bile acid is hindered, and the content of glycocholic acid in the reflux blood circulation increases. It also increases the content of glycocholic acid in the blood. Therefore, glycocholic...

Examples

preparation example Construction

[0096] In some embodiments, a preparation method of glycocholic acid detection kit is provided, comprising:

[0097] 1) Preparation of glycocholic acid-biotin

[0098] The preparation principle of glycocholic acid-biotin is as follows:

[0099]

[0100] Raw materials include glycocholic acid; NH 2 -PEG-Biotin is an amino PEG biotin derivative, the molecular weight of PEG can be 300-3000; condensing agents include but are not limited to N,N'-dicyclohexylcarbodiimide, 1-ethyl-(3-dimethyl Aminopropyl)carbodiimide hydrochloride, N,N'-diisopropylcarbodiimide, O-(7-azabenzotriazol-1-yl)-N,N,N', At least one of N'-tetramethylurea hexafluorophosphate and benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate; catalysts include but are not limited to 4-difluorophosphate At least one of methylaminopyridine, 1-hydroxybenzotriazole, and 4-pyrrolidinopyridine; solvents include but are not limited to dichloromethane, tetrahydrofuran, ether, ethyl acetate, dimethyl sulfoxide, N,N ...

Embodiment 1

[0113] In this example, the reagents used for pH adjustment were 2.0M hydrochloric acid and 2.0M sodium hydroxide aqueous solution.

[0114] In the present embodiment, the source of each raw material and product information are as follows:

[0115] Manufacturer of polystyrene carboxyl microspheres: JSR (Beijing Bormaier agent); product name: latex microspheres; product number: P0113.

[0116] Manufacturer of Glycocholic Acid: Aladdin; Product Name: Glycocholic Acid; Product Number: G131002.

[0117] Antibody: Manufacturer: Tansheng Technology; Product Name: CG Rabbit Polyclonal Antibody; Product Number: FAB-F005.

[0118] NH 2 -PEG-Biotin: Manufacturer: Shanghai Tuoyang; Product Name: Biotin Polyethylene Glycol Amino; Product Number: P002009.

[0119] 1. Preparation of glycocholic acid reagent

[0120] 1.1 Preparation of glycolic acid-biotin derivatives

[0121] To a dry round bottom flask was added glycocholic acid (46.5 mg, 0.10 mmol), NH 2-PEG-Biotin (PEG molecular we...