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Quantitative determination kit for proteinase 3 antibody (PR3-Ab) and detection method thereof

A quantitative measurement and kit technology, applied in the field of clinical immunology detection, can solve the problems of inability to distinguish non-specific recognition, expensive fluorescent microscope, and inability to carry out quantitative determination, etc., to achieve fast and reliable reaction process, high detection sensitivity and linear range , validity period and the effect of testing performance guarantee

Active Publication Date: 2014-10-08
北京贝尔医疗设备有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method cannot distinguish non-specific recognition according to the size of the molecular weight when analyzing the results; the operation is relatively complicated and requires a more expensive fluorescence microscope, which can only be used for qualitative detection and cannot be used for quantitative determination

Method used

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  • Quantitative determination kit for proteinase 3 antibody (PR3-Ab) and detection method thereof
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  • Quantitative determination kit for proteinase 3 antibody (PR3-Ab) and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, the preparation of magnetic separation reagent:

[0063] 1. The preparation process of magnetic particle buffer, taking the preparation of 1L as an example:

[0064] 1. Measure 800mL of purified water in a container, weigh 12.1g of Tris and 8.5g of NaCl into the container, stir well until completely dissolved;

[0065] 2. Weigh 5g of BSA, 50mL of newborn bovine serum, and 3000.2mL of Proclin into the container, stir well until completely dissolved;

[0066] 3. Use 4M HCl to adjust the pH, and control the pH between 7.9-8.1;

[0067] 4. Finally, dilute to 1L with purified water, filter with a 0.2um filter, and store at 2-8°C for later use;

[0068] 2. Preparation process of magnetic separation reagent

[0069] 1. Take magnetic particles containing carboxyl groups (COOH) active groups, the carboxyl content per gram (g) of magnetic particles (dry weight) is not less than 0.3 millimoles (mmoL), with superparamagnetic properties, and the diameter is between ...

Embodiment 2

[0078] Embodiment 2: the preparation of enzyme labeling reagent:

[0079] 1. The preparation process of enzyme labeling reagent diluent, taking the preparation of 1L as an example:

[0080] 1. Measure 800mL of purified water in a container, weigh 6.06g of Hepes and 8.5g of NaCl into the container, and stir until completely dissolved;

[0081] 2. Weigh BSA5g, Zncl 2 0.1g, Proclin-3000.2mL and MgCl 2 0.1g in the container, fully stirred until completely dissolved;

[0082] 3. Use 4M HCl to adjust the pH, and control the pH between 7.5-8.0;

[0083] 4. Finally, dilute to 1L with purified water and store at 2-8°C for later use;

[0084] 2. Coupling of alkaline phosphatase (ALP) to PR3 antibody

[0085] 1. PR3 antibody, purchased from Beijing Jiufeng Runda Biotechnology Co., Ltd., with a purity of more than 95%;

[0086] 2. The purity of alkaline phosphatase should exceed 95%, the specific activity should exceed 1000U / mg, and the concentration should exceed 5mg / mL;

[0087] ...

Embodiment 3

[0092] Embodiment 3, the preparation of calibrator:

[0093] 1. Preparation of calibrator diluent:

[0094] 1. Measure 600mL of purified water in a container, weigh 200mL of newborn bovine serum and add it to the container, stir well and mix evenly;

[0095] 2. Weigh 3.4g of dipotassium hydrogen phosphate, 0.36g of potassium dihydrogen phosphate, and 3000.02mL of Proclin into the container, stir well until completely dissolved;

[0096] 3. Use 4M HCl to adjust the pH, and control the pH between 7.0-7.5;

[0097] 4. Finally, dilute to 1L with purified water and store at 2-8°C until use.

[0098] 2. Preparation of calibrator:

[0099] 1. Prepare the PR3 antibody at 0, 5, 20, 50, 150, and 300 U / mL with the calibrator diluent, a total of 6 concentration points.

[0100]

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Abstract

Belonging to the technical field of clinical immunological detection, the invention discloses a quantitative determination kit for a proteinase 3 antibody (PR3-Ab). The kit includes a magnetic separation reagent, an enzyme labeled reagent, a calibrator, a cleaning concentrated solution, a substrate solution and a stabilizer. The invention also discloses a preparation method for the kit and a detection method for the kit. The kit prepared by the invention has the advantages of good accuracy, sensitivity and stability, low cost, simple operation, and broad application prospect.

Description

technical field [0001] The invention belongs to the technical field of clinical immunology detection, and in particular relates to a protease 3 antibody (PR3-Ab) quantitative assay kit and a detection method thereof. [0002] Background technique [0003] Protease 3 (PR3) is a serine protease in neutrophil cytoplasmic azurophilic granules, a glycoprotein with a molecular weight of about 29KDa. PR3 can degrade a variety of extracellular matrix such as elastin, hemoglobin, collagen type IV and other tissue components. Furthermore, proteinase 3 promotes platelet activation through cathepsin G and inactivates C1 inhibitors. PR3 antibody is a specific antibody for Weigla granulomatosis (WG). Although its pathogenesis is unclear, anti-PR3 antibodies have been implicated in the pathogenesis of VGD. In addition, the concentration of PR3 antibody is closely related to the activity of other diseases, which can be seen in primary systemic vasculitis, chronic inflammatory bowel dise...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N33/532
CPCG01N33/535G01N33/5375G01N33/573
Inventor 于大为杨晓勇
Owner 北京贝尔医疗设备有限公司
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