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Kit for detecting myeloperoxidase (MPO) concentration in sample and preparation method of kit

A technology of myeloperoxidase and horseradish peroxidase, which is applied in the field of kits for detecting the concentration of myeloperoxidase (MPO) in samples and its preparation, can solve the problem of high skill requirements for experimental operators and large disadvantages. Problems such as scope promotion and high detection cost, to achieve the effect of facilitating promotion, easy operation, and low detection cost

Inactive Publication Date: 2014-10-29
北京协和洛克生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used reagents for detecting MPO concentration on the market need to be equipped with expensive large-scale instruments. The detection cost is high, the operation is cumbersome, and the requirements for the skills of the experimental operators are high, which is not conducive to the large-scale promotion of this project.

Method used

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  • Kit for detecting myeloperoxidase (MPO) concentration in sample and preparation method of kit
  • Kit for detecting myeloperoxidase (MPO) concentration in sample and preparation method of kit
  • Kit for detecting myeloperoxidase (MPO) concentration in sample and preparation method of kit

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Effect test

Embodiment 1

[0034] A kit for detecting the concentration of myeloperoxidase in a sample, including a microwell plate coated with an anti-myeloperoxidase monoclonal antibody, a series of standard substances and quality controls for myeloperoxidase, horseradish peroxidase Anti-myeloperoxidase-labeled monoclonal antibody, sample buffer, hydrogen peroxide in citrate buffer (referred to as substrate A), tetramethylbenzidine in citrate buffer (referred to as substrate Substance B), 1-2M sulfuric acid solution (called stop solution), phosphate buffer solution containing surfactant (called concentrated washing solution).

[0035] Further, the microwell plate is a 96-well polystyrene microwell plate, which is coated with 1-50 μg / well of anti-myeloperoxidase monoclonal antibody.

[0036] Further, the standard myeloperoxidase series and the quality control are prepared by diluting the pure myeloperoxidase and the self-made standard diluent according to 1:200000-1:1000, and the standard The differen...

Embodiment 2

[0053] Prepare the kit by:

[0054] Preparation of microwell plates coated with anti-myeloperoxidase monoclonal antibody: Dilute the specific anti-myeloperoxidase monoclonal antibody with 20mM sodium phosphate buffer solution with a pH value of 7.4 to a concentration of 1 μg / ml , added to the concave well of a 96-well polystyrene microplate, 100ul / well, and coated overnight at 4°C; remove the coating solution, and then wash the microplate three times with a plate washer; Add 10mM sodium phosphate buffer solution of pH 7.4 containing 1% bovine serum albumin to the well: 300ul / well, seal at 37°C for 2 hours; shake off the blocking solution in the well, dry at 37°C for 4 hours, and seal in a vacuum bag , stored at 4°C;

[0055] Preparation of myeloperoxidase series standard and quality control products: use 20mM sodium phosphate buffer solution (pH7.4 ) Prepare pure myeloperoxidase into a series of standard products with marked concentrations of 0, 50, 100, 200, 400, 800ng / ml a...

Embodiment 3

[0075] Prepare the kit by:

[0076] Preparation of microwell plates coated with anti-myeloperoxidase monoclonal antibody: Dilute the specific anti-myeloperoxidase monoclonal antibody with 10 mM sodium phosphate buffer solution with a pH value of 8.0 to a concentration of 10 μg / ml , added to the concave well of a 96-well polystyrene microplate, 100ul / well, and coated overnight at 4°C; remove the coating solution, and then wash the microplate three times with a plate washer; Add 10mM sodium phosphate buffer solution at pH 8.0 containing 2% bovine serum albumin to the well: 300ul / well, seal at 37°C for 2 hours; shake off the blocking solution in the well, dry at 37°C for 4 hours, and seal in a vacuum bag , stored at 4°C;

[0077] Preparation of myeloperoxidase series standard and quality control products: Myeloperoxidase was mixed with 20mM sodium phosphate buffer solution (PH7.4) containing 2% goat serum, 0.1% sodium azide, 1% bovine gamma protein, and 0.1% Tween20. The pure p...

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Abstract

The invention belongs to the technical field of in-vitro immunodetection and in particular relates to a kit for detecting myeloperoxidase (MPO) concentration in a sample and a preparation method of the kit. The kit comprises a porous plate coated with an anti-MPO monoclonal antibody, an MPO series standard substance and a quality control material, a horse radish peroxidase-labeled anti-MPO monoclonal antibody, a sample buffer solution, a citrate buffer solution of hydrogen peroxide, a citrate buffer solution of tetramethyl benzidine, a 1-2M sulfuric acid solution, and a phosphate buffer solution containing surfactants. Because the used detection instruments are simple and are universal instruments, the kit is low in detection cost and contributes to popularization; the kit is easy to operate and can be realized without professionals. Meanwhile, according to the unique standard diluent and sample buffer solution formula in the kit, the reliability and the accuracy of the detection result are greatly improved.

Description

technical field [0001] The invention belongs to the technical field of in vitro immunoassay, and in particular relates to a kit for detecting the concentration of myeloperoxidase (MPO) in a sample and a preparation method thereof. Background technique [0002] Myeloperoxidase (MPO) is a leukocyte enzyme with a relative molecular mass of 150KD, which is a glycosylated heme protein. MPO originates from polymorphonuclear neutrophils, monocytes and macrophages, and is stored in azurophilic granules. When leukocytes are activated and degranulated, MPO is released into phagocytic vesicles and plasma. Under certain conditions, it can induce oxidative stress and tissue damage. As a marker of systemic inflammation and oxidative stress, MPO participates in the occurrence and development of coronary heart disease through various mechanisms. A large number of epidemiological studies have shown that the increase of MPO concentration is closely related to coronary heart disease, so detec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/573G01N33/545
CPCG01N33/723G01N33/545G01N33/573
Inventor 郑乐民王晓华吴建榕
Owner 北京协和洛克生物技术有限责任公司
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