Human myeloperoxidase chemiluminescent immunodetection kit

A myeloperoxidase and chemiluminescence immunological technology, applied in the field of immunoassay medicine, to achieve the effects of good accuracy, improved sensitivity and high stability

Inactive Publication Date: 2013-04-10
天津市协和医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Currently, chemiluminescence immunoassay technology is s

Method used

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  • Human myeloperoxidase chemiluminescent immunodetection kit
  • Human myeloperoxidase chemiluminescent immunodetection kit
  • Human myeloperoxidase chemiluminescent immunodetection kit

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0031] Example 1

[0032] A human myeloperoxidase chemiluminescent immunoassay kit, comprising the following components:

[0033] 1) Human myeloperoxidase standard;

[0034] 2) Microwell plate coated with human myeloperoxidase monoclonal antibody;

[0035] 3) Alkaline phosphatase-labeled human myeloperoxidase monoclonal antibody;

[0036] 4) The chemiluminescent substrate AMPPD that the above enzyme acts on;

[0037] 5) Concentrated washing liquid;

[0038] 6) Composition of semi-finished products and finished products;

[0039] in:

[0040] 1. The human myeloperoxidase standard product is prepared by the following method: mix 0.02mol / L phosphate buffer solution with a pH value of 7.4 and fetal bovine serum in a volume ratio of 4:1 to prepare a basic buffer solution, and use Human myeloperoxidase was diluted to concentrations of 0, 25, 64, 160, 400, and 1000 ng / mL in basic buffer solution.

[0041] 2. The preparation steps of the microwell plate coated with human myel...

Example Embodiment

[0066] Example 2

[0067] A human myeloperoxidase chemiluminescent immunoassay kit, comprising the following components:

[0068] 1. Human myeloperoxidase standard substance (same as Example 1)

[0069] 2. Microwell plate coated with human myeloperoxidase monoclonal antibody (same as Example 1)

[0070] 3. Horseradish peroxidase-conjugated monoclonal antibody to human myeloperoxidase

[0071] Dissolve 4 mg of horseradish peroxidase (HRP) in 1 mL of deionized water, add 0.4 mL of 50 mM sodium periodate aqueous solution, shake at room temperature for 30 min, dialyze overnight with 1 mM sodium acetate buffer (pH 4.4), add 6 mg of MPO monoclonal antibody, shake overnight at 2-8°C, reduce with 400 μL of 200 mM NaBH4, then dialyze through 0.02M PBS with a pH value of 7.4 overnight, use HPLC for secondary purification, collect protein peaks and add an equal volume Glycerol, stored at -20°C.

[0072] 4. The chemiluminescent substrate luminol acted by the above enzymes

[0073] Th...

Example Embodiment

[0088] Embodiment 3 The using method of kit of the present invention

[0089] The use of the human MPO chemiluminescent immunoassay kit prepared in Example 1, the specific operations are as follows:

[0090] 1) Take out the kit of the present invention from the refrigerator at 4°C, and equilibrate to room temperature;

[0091] 2) Take a microwell plate prepared in Step 2 of Example 1 and number it, and repeat all experiments with double holes;

[0092] 3) Add 25 μL each of the calibrator and the sample to be tested at a concentration of 0, 25, 64, 160, 400, and 1000 ng / mL into the wells of the microplate, and set a blank well for each experiment, and then remove the blank well Add 100 μL of the enzyme-labeled human myeloperoxidase monoclonal antibody prepared in Step 3 of Example 1 to each well;

[0093] 4) Oscillate on the oscillator for 30s to mix;

[0094] 5) Seal the plate with parafilm and incubate at 37°C for 45 minutes;

[0095] 6) Shake off the reaction solution, w...

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Abstract

The invention discloses a human myeloperoxidase (MPO) chemiluminescent immunodetection kit. The kit comprises the following components: 1) a human MPO standard product; 2) a vector coating a human MPO monoclonal antibody; 3) the enzyme-labeled human MPO monoclonal antibody; 4) a chemiluminescent substrate on which the enzyme acts; and 5) a concentrated cleaning solution. The kit can perform concentrated quantitative detection on the MPO molecules in a serum sample of a patient. The kit effectively utilizes a chemiluminescent technology principle and adopts the enzyme to catalyze the chemiluminescent substrate on the basis of enzyme-linked immunoassay so as to improve the detection sensitivity, has the advantages of high stability, high specificity, excellent accuracy, simplicity in operation and no radioactive pollution, and can provide specific, quick and reliable basis for clinical diagnosis of cardiovascular diseases such as atherosclerosis (AS), acute coronary syndrome (ACS) and coronary heart disease (CHD).

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, specifically, the invention provides a human myeloperoxidase chemiluminescent immunoassay kit. Background technique [0002] Cardiovascular disease (CVD) is a high-risk disease that seriously endangers human life and health. At present, it has become the number one cause of death of the urban and rural population in my country, and it is also the non-communicable disease that causes the highest mortality rate. WHO pointed out that early diagnosis and prevention of CVD can effectively reduce the incidence of clinical emergencies and premature death. [0003] In recent years, a variety of cardiovascular disease markers have come out one after another. Since most of the indicators that can reflect myocardial necrosis and injury are detected after substantial myocardial damage occurs, it is difficult to provide a scientific basis for prevention in the early stage of the disease. Myeloperoxida...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
Inventor 单存海潘学继王立凯曹青洪宇霞
Owner 天津市协和医药科技集团有限公司
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