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Combination comprising thrombin agent and examining agent

A technology of thrombin and antithrombin, which is applied in the field of stable thrombin reagents and its inspection reagent kits, can solve the problems that the thrombin stabilization method is not perfect and does not show stability, etc.

Inactive Publication Date: 2004-06-16
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] However, none of these is perfect as a method for stabilizing thrombin, and it has not shown sufficient stability especially as a diagnostic drug for clinical examination.

Method used

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  • Combination comprising thrombin agent and examining agent
  • Combination comprising thrombin agent and examining agent
  • Combination comprising thrombin agent and examining agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Benzamidine chlorate was added to the thrombin reagent (manufactured by International Reagent Co., Ltd.) at the following concentration. This thrombin reagent is a solution made of 200U / mL thrombin, acetic acid and a buffer containing calcium lactate, with a pH value of 6.0.

[0079] No.1: Thrombin solution (without adding benzamidine).

[0080] No.2: Add benzamidine to No.1 to make 5 mM benzamidine chlorate.

[0081] No.3: Add benzamidine to No.1 to make 10 mM benzamidine chlorate.

[0082] The residual activity of each thrombin was measured when the above samples were stored at 37°C for 0, 1, 2, 4 and 8 weeks. The determination method is in accordance with the method stipulated by the Japanese Pharmacopoeia. Measurement results such as figure 1 Shown: after 8 weeks, the thrombin activity of the group without benzamidine chlorate (No.1) dropped to below 50%; The activity residue is about 75% or more.

Embodiment 2

[0084] The thrombin solution described in No.1 of Example 1 is diluted with a buffer as shown in Example 1 to prepare a solution with a thrombin activity of 100 U / mL, and add benzamidine chlorate to make it reach the following concentration, and the thrombin stabilization effect at this time was investigated.

[0085] No.4: Thrombin solution (100U / mL, no benzamidine added)

[0086] No.5: Add benzamidine to No.4 to make 2.5 mM benzamidine chlorate.

[0087] No.6: Add benzamidine to No.4 to make 5 mM benzamidine chlorate.

[0088] Regarding the above-mentioned samples, the residual thrombin activity of each sample was measured in the same manner as in Example 1. Measurement results such as figure 2 Shown: In the group without benzamidine chlorate (No.4), the thrombin activity dropped to below 50% after 8 weeks; in the group added with benzamidine chlorate, the thrombin activity remained above 75%.

Embodiment 3

[0090] The thrombin solution described in No.1 of Example 1 was diluted with buffer as shown in Example 1 to prepare solutions with thrombin activities of 100U / mL, 120U / mL, 150U / mL and 200U / mL, Benzamidine was added separately so that the concentrations of benzamidine chlorate were 0, 2.5mM, 5.0mM, 7.5mM, 10.0mM, 12.5mM, and 15.0mM, and they were stored at 37°C for 8 weeks. Thrombin stabilization effect.

[0091] Measurement results such as image 3 Shown: In the solutions containing thrombin at various concentrations, the thrombin activity dropped below 50% in the group without adding benzamidine chlorate; in the group adding 2.5mM benzamidine chlorate, the thrombin activity remained at 70% % or more; in the group added with 7.5mM, the thrombin activity remained above 80%.

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Abstract

Provided is a stabilized thrombin containing a test reagent and the test reagent kit. The test reagent contains thrombin and an inhibitor having thrombin inhibiting effect, the inhibitor is a protease antagonist having thrombin inhibiting effect, and is preferably selected from the group consisting of benzamidine, p-aminobenzamidine, m-aminobenzamidine, phenylguanidine, argatroban, gabexate mesylate and nafamostat mesylate. The test reagent of the present invention can contain one or more compounds with thrombin stabilizing function, selected from the group consisting of a calcium ion, an organic acid, a surfactant and a protein. The test reagent is useful as a solidifying ability test reagent, especially, fibrinogen metering reagent.

Description

technical field [0001] The present invention relates to a stable thrombin-containing composition (thrombin reagent) and a testing reagent set thereof. More specifically, it relates to a stable thrombin reagent and a test reagent set thereof used in clinical medicine, especially in the field of fibrinogen quantification. Background technique [0002] The enzyme thrombin, which converts fibrinogen into fibrin, plays a very important role in the blood clotting reaction process. In the field of clinical examination, thrombin is widely used to measure the concentration of fibrinogen, the activity of thrombin-inhibiting substances, such as antithrombin activity and hirudin activity, and to further determine the activity of chemical synthesis inhibitors. [0003] However, since thrombin is generally not stable, when long-term storage is required, it is usually lyophilized before storage. In addition, to date, various attempts have been made to stabilize thrombin, as described in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/56
Inventor 菊川纪弘
Owner SYSMEX CORP
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