Iron chelator-containing prothrombin time reagent

A technology of prothrombin time and iron chelating agent, which is applied in the field of coagulation diagnosis and can solve problems such as poor stability

Active Publication Date: 2020-07-10
德国西门子医学诊断产品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a problem with providing prothrombin time reagents in liquid form is their poor stability

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Determination of the Stability of Liquid Prothrombin Time Reagent Stabilized by Desferrioxamine

[0038] at time point t 0 , deferoxamine (deferoxamine mesylate, Sigma-Aldrich Chemie GmbH, Steinheim, Germany) was added in different batches to liquid coagulation containing recombinant human tissue factor protein, synthetic phospholipids and calcium ions at the following final concentrations In the zymogen time reagent:

[0039] 1.0 mM deferoxamine,

[0040] 2.0.0137mM (9mg / L) deferoxamine,

[0041] 3.0.0183mM (12mg / L) deferoxamine,

[0042] 4.0.0228mM (15mg / L) deferoxamine,

[0043] 5.0.114mM (75mg / L) deferoxamine,

[0044] 6. 0.228mM (150mg / L) deferoxamine.

[0045] exist Different reagents were used in the automated prothrombin time test (PT test) in the CA-1500 analyzer (Sysmex Corp., Kobe, Japan). Use the control plasma specified below as a sample:

[0046] · Clotting Control Level 1 (Ci-Trol 1) control plasma; normal control used to determine ...

Embodiment 2

[0052] Example 2: Determination of the stability of liquid prothrombin time reagents stabilized with different iron chelators

[0053] at time point t 0 In different batches, the following metal ion chelating agents were added to the liquid prothrombin time reagent containing recombinant human tissue factor protein, synthetic phospholipids and calcium ions at 0.0228mol / L:

[0054] i. Fluorescent siderophores (Pyoverdines, Sigma-Aldrich Chemie GmbH, Steinheim, Germany);

[0055] ii. Iron pigment (free iron iron pigment from Ustilago sphaerogena, Sigma-Aldrich Chemie GmbH, Steinheim, Germany);

[0056]iii. deferoxamine (deferoxamine mesylate, Sigma-Aldrich Chemie GmbH, Steinheim, Germany); deferoxamine (deferoxamine mesylate, Sigma-Aldrich Chemie GmbH, Steinheim, Germany) );

[0057] iv. Deerasirox (Combi-Blocks, Inc., San Diego, USA);

[0058] v.DTPA (diethylenetriaminepentaacetic acid);

[0059] vi.EDTA (ethylenediaminetetraacetic acid);

[0060] vii. BAPTA (1,2-bis(2-am...

Embodiment 3

[0069] Example 3: Determination of the presence of higher Fe 3+ Stability of liquid prothrombin time reagents stabilized with different iron chelators at ionic concentrations

[0070] at time point t 0 , the FeCl of 0.00308mM (0.5mg / L) 3 Added to the liquid prothrombin time reagent containing recombinant human tissue factor protein, synthetic phospholipids and calcium ions, and then added 0.0228mM of the metal ion chelating agent mentioned in Example 2 in different batches.

[0071] As described in Example 1, in Different reagents were used in the automated prothrombin time test (PT test) in the CA-1500 analyzer (Sysmex Corp., Kobe, Japan). Quality control plasma Ci-Trol 1 and Ci-Trol 2 described in detail in Example 1 were used as samples.

[0072] In order to determine the presence of higher Fe 3+ Long-term stability of the different reagents at ion concentration, the reagents were stored in liquid state at +37°C in stoppered glass vials over a period of 6 weeks. Reag...

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Abstract

The present invention pertains to the field of coagulation diagnostics and relates to a prothrombin time reagent on the basis of recombinant or native tissue factor protein and phospholipids, which reagent can be stabilized by the addition of an iron chelator from the groups of siderophores and 3,5-diphenyl-1,2,4-triazoles.

Description

technical field [0001] The invention belongs to the field of coagulation diagnosis, and relates to prothrombin time reagents based on recombinant or natural tissue factor protein and phospholipids, which can be selected from siderophore and 3,5-diphenyl-1,2 by adding , 4-triazoles in the group consisting of iron chelating agents to stabilize. Background technique [0002] Tissue factor protein (abbreviated as tissue factor or thromboplastin, tissue factor in English) is a transmembrane protein that is crucial for blood coagulation. It is expressed by cells that are not normally in contact with flowing blood, for example by cells in the subendothelium (smooth muscle) and perivascular cells such as fibroblasts. However, in the case of damaged blood vessels, cells expressing the tissue factor protein come into contact with factor VII, a proclotting factor that circulates in the blood. In the presence of calcium, tissue factor protein and Factor VII form a complex, and the act...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86G01N2333/7454G01N2800/224
Inventor 萨比内·皮尔格林托马斯·维塞尔诺贝特·赞德
Owner 德国西门子医学诊断产品有限公司
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