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Method of stabilizing thrombin and composition thereof

A technology of thrombin and composition, applied in the direction of biochemical equipment and methods, enzymes, peptidases, etc., can solve problems such as thrombin instability and reduced blood coagulation activity

Inactive Publication Date: 2017-10-13
VITRUM BIOLOGIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, thrombin is inherently unstable and its activity for blood clotting is significantly reduced even after only 2-3 hours after fresh preparation

Method used

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Embodiment Construction

[0011] The present invention is based on the unexpected discovery that thrombin can be stabilized via heat treatment.

[0012] In one aspect, the present invention provides a method of stabilizing thrombin in a thrombin solution comprising heating the thrombin solution at a temperature of 35-85° C. for 1 to 20 seconds, and quenching the heating.

[0013] The thrombin solution can be prepared from blood by methods known in the art. In one example, 6-7 mL of thrombin solution can be prepared from about 10 mL of platelet poor plasma (PPP) by a thrombin generating device.

[0014] According to a particular embodiment, the heating is performed in a water bath. As an example, 1.5 mL of thrombin solution in a microcentrifuge tube was placed in a 35-85°C water bath for 1 to 20 seconds, then quenched in ice water to obtain a stabilized thrombin solution.

[0015] Those skilled in the art can determine the appropriate combination of heating temperature and time. For example, higher h...

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Abstract

A method of stabilizing thrombin in a thrombin solution is provided. The method comprises heating the thrombin solution at a temperature of 35-85 DEG Cfor 1 to 20 seconds, and quenching the heating. Also provided is a stabilized thrombin composition. The stabilized thrombin composition is characterized in that it is prepared by heating a thrombin solution at a temperature of 35-85 DEG C for 1 to 20 seconds, and quenching the heating. The thrombin solution may be further lyophilized to obtain a stabilized thrombin composition in lyophilized form.

Description

technical field [0001] The present invention relates to a method of stabilizing thrombin in a thrombin solution. Background technique [0002] Thrombin is a serine protease that plays a key role in the process of blood coagulation. In the coagulation mechanism, prothrombin is cleaved to form thrombin, which ultimately results in a reduction in blood loss. Thrombin then acts as a serine protease that converts soluble fibrinogen into insoluble fibrin, which in turn promotes blood clotting. [0003] However, thrombin is inherently unstable and its activity for blood clotting is markedly reduced even after only 2-3 hours after being freshly prepared. In the past, there have been extensive attempts on how to stabilize thrombin, but they were basically chemical methods. For example, EP 1221479B1 discloses the use of a nonionic surfactant and calcium ions, while US 8071090B2 discloses the use of a preservative of benzyl alcohol or chlorobutanol and sucrose. [0004] Given that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/48
CPCC12N9/6429C12Y304/21005
Inventor 苏正尧孙崇谨
Owner VITRUM BIOLOGIES
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