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Myeloperoxidase detection reagent and preparation and use method thereof

A technology of myeloperoxidase and myeloperoxidase, applied in the field of myeloperoxidase detection reagents

Active Publication Date: 2019-06-14
苏州博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a lack of myeloperoxidase detection reagents with good stability, high sensitivity and strong specificity in the market, especially good quality automatic detection reagents

Method used

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  • Myeloperoxidase detection reagent and preparation and use method thereof
  • Myeloperoxidase detection reagent and preparation and use method thereof
  • Myeloperoxidase detection reagent and preparation and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Synthesis of myeloperoxidase amino-terminal tripeptide derivatives

[0057] The chemical structural formula of the tripeptide derivative at the amino terminal of myeloperoxidase is shown in formula (Ⅲ):

[0058]

[0059] Formula (Ⅲ).

[0060] The synthesis route and preparation steps of the above-mentioned myeloperoxidase amino-terminal tripeptide derivatives are as follows:

[0061]

[0062] Concrete synthetic steps are as follows:

[0063] (I) Synthesis of compound 3: Dissolve 22 g of compound 1 in 100 ml of dimethylformamide (DMF), then add 32 g of tetramethylurea tetrafluoroborate (TBTU), 10 g of compound 2 and 20 g of Triethylamine (TEA) was used to make the reaction mixture, and the above mixture was stirred at room temperature for 2 hours, then poured into 200 ml of purified water, filtered, and the filter cake was dried in vacuo to obtain 24 g of compound 3 as a gray solid, yield 79%.

[0064]

[0065] (II) Synthesis of compound 4: Add 24...

Embodiment 2

[0078] Example 2. Preparation of myeloperoxidase amino-terminal tripeptide immunogen

[0079] The myeloperoxidase amino-terminal tripeptide immunogen in this example is formed by linking the myeloperoxidase amino-terminal tripeptide derivative represented by formula (III) with bovine serum albumin (BSA), and its structural formula is as follows Formula (I) shows:

[0080]

[0081] Formula (Ⅰ).

[0082] The specific steps of the synthesis method of the myeloperoxidase amino-terminal tripeptide immunogen are as follows:

[0083] (1) Dissolve the carrier protein with a mass fraction of 1.0% in 0.2 mmol / L, pH=8.5 phosphate buffer to obtain a carrier protein solution;

[0084] (2) Dissolve 0.5% of myeloperoxidase amino-terminal tripeptide derivatives, 5.0% of dimethylformamide, and 5.0% of ethanol in 10 mmol / L, pH=5.0 potassium phosphate buffer , then add 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide with a mass fraction of 0.5%, and stir and react the above chemical substanc...

Embodiment 3

[0086] Example 3. Preparation of anti-myeloperoxidase specific antibody

[0087] The myeloperoxidase amino-terminal tripeptide immunogen prepared in Example 2 was inoculated into experimental animal rabbits by conventional methods, and the antiserum was taken after booster immunization. The specific steps were as follows:

[0088] a. Dilute the myeloperoxidase amino-terminal tripeptide immunogen to 3.0 mg / ml with PBS buffer to obtain an immunogen solution, and then use 3.0 ml of the immunogen solution to mix with an equal amount of Freund's complete adjuvant. Experimental animal rabbits were injected;

[0089] b. After 2 weeks, mix 3.0 ml of the same immunogen solution with the same amount of Freund's incomplete adjuvant, inject once into the above-mentioned experimental animal rabbit, and then inject once every 3 weeks, a total of 5 injections;

[0090] c. Take blood from the immunized experimental animal rabbit, separate and purify to obtain specific anti-myeloperoxidase an...

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Abstract

The invention relates to a myeloperoxidase detection reagent and a preparation and use method thereof. The automatic determination of the myeloperoxidase content on a full-automatic biochemical analyzer can be realized by applying the detection reagent, the myeloperoxidase content in a biological sample can be quickly and precisely determined in high flux, and the detection reagent has the advantages of being simple in operation, high in sensitivity, strong in specificity, and accurate in result. the myeloperoxidase detection cost can be effectively reduced, and the detection reagent is in favor of clinical and extensive popularization and use.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a myeloperoxidase detection reagent and a preparation and use method thereof. Background technique [0002] Myeloperoxidase (Myeloperoxidase, MPO) is a hemoglobin protein that exists in the azurophilic granules of neutrophil multinucleated cells and macrophages, and its main function is to use hydrogen peroxide and chloride ions to produce hypochlorous acid Salt, and form free radicals with oxidizing ability to kill microorganisms in phagocytes. Moreover, MPO can be released outside the cells to destroy a variety of target substances, such as tumor cells, platelets, NK cells, protozoa, toxins, etc., and play a role in the production and regulation of inflammatory responses in the body. Therefore, MPO content is a sensitive indicator in inflammatory lesions. [0003] At present, the methods used to detect the content of myeloperoxidase at home and abroad...

Claims

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Application Information

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IPC IPC(8): G01N33/72G01N33/573G01N33/531
Inventor 虞留明
Owner 苏州博源医疗科技有限公司
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