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Thrombomodulin chemiluminescence quantitative immunoassay detection kit and preparation method thereof

A technology of thrombomodulin and chemiluminescence immunity, which is applied in the field of immunodiagnosis, can solve the problems of complicated operation, unstable test results, and low accuracy, and achieve the effects of simple operation, improved specificity and detection speed, and low cost

Inactive Publication Date: 2019-09-03
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the technical problems of low precision, complex operation and unstable test results of the thrombomodulin method in the prior art, and provide a thrombomodulin chemiluminescent immunoquantitative detection kit and its preparation method

Method used

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  • Thrombomodulin chemiluminescence quantitative immunoassay detection kit and preparation method thereof
  • Thrombomodulin chemiluminescence quantitative immunoassay detection kit and preparation method thereof
  • Thrombomodulin chemiluminescence quantitative immunoassay detection kit and preparation method thereof

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preparation example Construction

[0050] The present invention also provides a preparation method of the thrombomodulin chemiluminescent immunoquantitative detection kit, comprising the following steps:

[0051] Step 1, preparation of R1 reagent

[0052] Add the streptavidin magnetic particle solution into the TBST solution, mix well, place on the magnetic separator until the supernatant is free of turbidity, discard the supernatant, keep the magnetic particles, wash the magnetic particles with buffer Ⅰ, Then use buffer I to make a solid-phase reagent with a magnetic particle concentration of 0.007% to 1%, which is the R1 reagent;

[0053] Among them, the ratio of streptavidin magnetic particles to TBST solution is preferably 50mg:10mL; general cleaning needs to be repeated three times; R1 reagent needs to be stored at 2-8°C.

[0054] Step 2, preparation of R2 reagent

[0055] First put the thrombomodulin monoclonal antibody into the centrifuge tube to ensure that the thrombomodulin monoclonal antibody is at...

Embodiment 1

[0070] The preparation method of the chemiluminescent immunoquantitative detection kit of thrombomodulin comprises the following steps:

[0071] Preparation of R1 reagent: Take 0.5 ml (50 mg) of a solution of streptavidin magnetic particles (particle size 0.1 μm) with a concentration of 100 mg / ml and add it to the TBST solution, mix well, and place it on a magnetic separator until the supernatant If there is no turbidity, discard the supernatant, keep the magnetic particles, and wash them three times with buffer I. After washing, use buffer I to make a solid-phase reagent with a magnetic particle concentration of 0.05%, which is the R1 reagent, and store at 2-8°C. ;

[0072] Preparation of R2 reagent: put 500 μg of thrombomodulin monoclonal antibody into a centrifuge tube, ensure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20 seconds), add carbonic acid buffer solution, mix well, add 5 μl 2mg / mL after mixing Centrifuge the DMF...

Embodiment 2

[0076] Preparation of R1 reagent: Take 0.5 ml (50 mg) of a solution of streptavidin magnetic particles (particle size 0.5 μm) with a concentration of 100 mg / ml and add it to the TBST solution, mix well, and place it on a magnetic separator until the supernatant If there is no turbidity, discard the supernatant, keep the magnetic particles, and wash them three times with buffer I. After washing, use buffer I to make a solid-phase reagent with a magnetic particle concentration of 1%, which is the R1 reagent, and store at 2-8°C. ;

[0077] Preparation of R2 reagent: put 500 μg of thrombomodulin monoclonal antibody into a centrifuge tube, ensure that the antibody is at the bottom of the centrifuge tube (centrifuge at room temperature for 20 seconds), add carbonic acid buffer solution, mix well, add 5 μl 2mg / mL after mixing Centrifuge the DMF solution of acridinium ester at room temperature for 0.5 min; seal the centrifuge tube with parafilm and put it in a light-proof cassette, th...

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Abstract

The invention relates to a thrombomodulin chemiluminescence quantitative immunoassay detection kit and a preparation method thereof, and belongs to the technical field of immunodiagnosis. The technical problem that the thrombomodulin method is low in accuracy, or complex in operation and unstable in test result in the prior art is solved. The thrombomodulin chemiluminescence quantitative immunoassay detection kit provided by the invention comprises an R1 reagent, an R2 reagent and an R3 reagent, wherein the R1 reagent comprises streptavidin magnetic particles and a buffer solution I, the R2 reagent comprises a thrombomodulin monoclonal antibody labeled by a chemiluminiscence marker and the buffer solution I, and the R3 reagent comprises a conjugation labeled thrombomodulin monoclonal antibody and the buffer solution I. The kit detects thrombomodulin by using a chemiluminescence immunoassay method, and has the advantages of high detection sensitivity, accurate quantitative detection, simple operation, no radioactivity risk, short detection time, and capability of detecting antigen and antibody target substances.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and particularly relates to a thrombomodulin chemiluminescence immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Thrombomodulin (Thrombomodulin, TM) is a single-chain glycoprotein with a relative molecular mass of 75,000 and a relative molecular mass of 105,000 after degrading disulfide bonds. It is synthesized by megakaryocytes and endothelial cells, widely distributed on the surface of vascular endothelial cells, and also exists in placental trophoblast cells, platelets, megakaryocytes, monocytes, neutrophils, synovial cells, keratinocytes, meningeal cells, smooth muscle cells, tumor cells. TM is a cofactor for thrombin-activated protein C. Combined with thrombin, it can reduce the coagulation activity of thrombin and enhance its activity of activating protein C. Since activated protein C has anticoagulant effect, TM is an important intr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/532
CPCG01N33/532G01N33/54326G01N33/6803
Inventor 高智玲翟涛高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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