Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Thrombin-antithrombin III compound determination kit as well as preparation method and application thereof

An antithrombin and thrombin technology, applied in the field of medical diagnosis, to achieve the effects of high sensitivity, high cost performance and good stability

Pending Publication Date: 2022-07-05
GUANGZHOU WONDFO BIOTECH
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is no mature method for the detection of thrombin-antithrombin III complex by chemiluminescent technology in China.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Thrombin-antithrombin III compound determination kit as well as preparation method and application thereof
  • Thrombin-antithrombin III compound determination kit as well as preparation method and application thereof
  • Thrombin-antithrombin III compound determination kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] This embodiment provides a thrombin-antithrombin III complex assay kit, the kit includes (1) a solution of a magnetic microsphere conjugate coated with a thrombin monoclonal antibody; (2) labeled with a thrombin monoclonal antibody Solution of enzyme-labeled conjugate of antithrombin III monoclonal antibody; (3) substrate solution; (4) calibrator.

[0062] The magnetic microsphere conjugate coated with thrombin monoclonal antibody is a combination of magnetic particle and thrombin monoclonal antibody; the diameter of the magnetic particle is 1000 nanometers to 3000 nanometers; In the solution of the magnetic microsphere conjugate of the monoclonal antibody, the concentration of the magnetic microsphere conjugate coated with the thrombin monoclonal antibody was 0.2 mg / mL. The concentration of thrombin monoclonal antibody in the magnetic microsphere conjugate was 4 μg / mL.

[0063] In the solution of the enzyme-labeled conjugate labeled with antithrombin III monoclonal an...

Embodiment 2

[0084] Example 2 Comparison of antigen coupling conditions

[0085] The changes in this example from Example 1 include the comparison of the preparation conditions of the thrombin-antithrombin III complex:

[0086] (1) Selection of crosslinking agent:

[0087] Three experimental groups were set up, each group took 10 mg of thrombin to replace the stock solution with PBS buffer, added 1 mg of Sulfo-EGS, Sulfo-MBS and DSS respectively, incubated for 1 h at room temperature, and activated to obtain a mixture; The original solution of thrombin III was replaced with PBS (phosphate) buffer, and all of them were immediately added to the above three groups of mixed solutions, and incubated for 1 h at room temperature for cross-linking. After the cross-linking was completed, desalted and purified, and mixed to obtain 10 mL of thrombin-antithrombin III complex solution each.

[0088] (2) Comparison of thrombin and Sulfo-EGS mass ratio conditions:

[0089]Three experimental groups wer...

Embodiment 3

[0097] Example 3: Performance evaluation of the kit of the present invention

[0098] 1. Standard curve

[0099] The thrombin-antithrombin III complex described in Example 1 was diluted with the standard substance diluent to prepare calibrator solutions S0-S9 of different concentrations, the concentrations were 0, 0.5ng / mL, 1.0ng / mL, 2.0 ng / mL, 4.0ng / mL, 9.0ng / mL, 18ng / mL, 36ng / mL, 60ng / mL, 120ng / mL, and stored at -20°C for later use. Then, use the kit provided in Example 1 of the present invention to detect the calibrators, and read the luminescence intensity values ​​corresponding to each calibrator respectively. The standard curve was obtained by fitting the concentration as the abscissa and the luminescence intensity as the ordinate.

[0100] 2. Linear

[0101] The basal serum sample (concentration≤blank limit) or calibrator dilution (as much as possible to use real high-value serum samples) with added antigens is used as the high-concentration sample, and the basal ser...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Concentrationaaaaaaaaaa
Login to View More

Abstract

The invention relates to a thrombin-antithrombin III compound antigen compound, a preparation method thereof and a thrombin-antithrombin III compound determination kit, and the thrombin-antithrombin III compound is prepared by cross-linking thrombin with a cross-linking agent Sulfo-EGS and then cross-linking with antithrombin III. The thrombin-antithrombin III compound determination kit is a chemiluminescence method, takes a thrombin-antithrombin III compound as a proofreading product, and has the advantages of high sensitivity, good stability and strong specificity, and is not easy to interfere. According to the detection method of the thrombin-antithrombin III compound, the probabilities that the dosage of a detection sample is small and the detection is faster are achieved, and a better choice is provided for clinical TAT quantification.

Description

technical field [0001] The invention belongs to the field of medical diagnosis, in particular to a thrombin-antithrombin III complex enzymatic chemiluminescence assay kit and a preparation method and application thereof. Background technique [0002] The hallmark of blood clotting is the formation of thrombin. Thrombin can promote the conversion of fibrinogen in plasma into insoluble fibrin, which is closely related to the formation of thrombus. However, thrombin produced in humans has a short half-life, making direct measurement of thrombin very difficult. After prothrombin is activated, it can quickly covalently link with antithrombin III to form thrombin-antithrombin III complex (TAT), and its half-life is long (10-30min), so the concentration of TAT can be determined by measuring the concentration of TAT. to reflect the thrombin concentration. [0003] In general cases, prothrombin is converted into thrombin, which is quickly neutralized by antithrombin III. Direct me...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/86G01N33/573G01N33/577G01N33/535G01N33/543G01N21/76
CPCG01N33/86G01N33/573G01N33/577G01N33/535G01N33/54326G01N21/76G01N2333/974
Inventor 杨建吴晗琪黄金梅王保升
Owner GUANGZHOU WONDFO BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products