Preparation method of porcine thrombin

A technology for porcine thrombin and prothrombin, which is applied in the field of thrombin preparation, can solve the problems of low yield of porcine thrombin, unsuitable for large-scale production, etc., and achieves improved biological safety, convenient large-scale production, and high thrombin harvest. rate effect

Inactive Publication Date: 2013-06-19
黑龙江迪龙制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the problems of low yield and unsuitability for large-sca...

Method used

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Experimental program
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Effect test

specific Embodiment approach 1

[0019] Specific embodiment one: the preparation method of a kind of porcine thrombin of the present embodiment is carried out according to the following steps:

[0020] 1. Separation of pig plasma: take fresh pig blood, add anticoagulant, and centrifuge to obtain pig plasma; wherein, the mass ratio of fresh pig blood to anticoagulant is 250:1;

[0021] 2. Chemical virus inactivation: add virus inactivation reagent according to 10% of the total weight of pig plasma, and incubate at 24-26°C for 6-8 hours to inactivate virus;

[0022] 3. Adsorption of prothrombin: add ion exchange resin according to 1.5% of the total weight of porcine plasma, adsorb at room temperature for 40-60 minutes, then filter and collect ion exchange resin with filter cloth, and use 5-10 times the washing liquid of ion exchange resin weight , washing the ion exchange resin, discarding the washing liquid;

[0023] 4. Collect prothrombin: the ion exchange resin washed with the washing solution in step 3 is ...

specific Embodiment approach 2

[0028] Embodiment 2: This embodiment differs from Embodiment 1 in that the anticoagulant described in step 1 is trisodium citrate with a mass percentage of 0.4%. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0029] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the virus inactivation reagent described in step two comprises the aqueous solution of polysorbate-80 and tributyl phosphate, wherein polysorbate methyl Ester-80 is 11% by volume, tributyl phosphate is 3.3% by volume, and the rest is water. Others are the same as in the first or second embodiment.

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Abstract

A preparation method of porcine thrombin relates to a preparation method of thrombin. The preparation method of porcine thrombin is used for solving the problems of low yield and unsuitability for large-scale production of the existing preparation method of porcine thrombin. The method provided by the invention comprises the following steps of: 1, separating porcine plasma; 2, inactivating virus by a chemical method; 3, absorbing prothrombin; 4, collecting the prothrombin; 4, nano-filtering; 6, activating the prothrombin; and 7, freeze-drying and storing the prothrombin. The preparation method provided by the invention is capable of improving the biosecurity of the thrombin product, and simultaneously, obtains higher thrombin yield rate by employing a special ion exchange resin extraction process; specifically, 80000 IU/L plasma can be obtained. According to the invention, the method of directly and proportionally feeding the ion exchange resin into the plasma is adopted, one ton to several tons of plasma can be processed each time; therefore, large-scale production is realized. The preparation method of porcine thrombin is applied to the industrial production field of thrombin.

Description

technical field [0001] The invention relates to a preparation method of thrombin. Background technique [0002] Thrombin is a serine protease formed after activation of a thrombin precursor (prothrombin) related to the blood coagulation system that widely exists in the blood of animals and humans. enzymes. [0003] Clinically, thrombin is suitable for ligation of small blood vessels and capillaries that are difficult to stop bleeding, and hemostasis of bleeding from solid organs. Hemostatic thrombin used in trauma, surgery, oral cavity, ENT, urology, burns, orthopedics, and other bleeding can directly act on fibrinogen in the blood, prompting the transformation into fibrin, accelerating blood coagulation and stopping bleeding. It is clinically used for hemostasis in trauma, surgery, oral cavity, otolaryngology, urology, obstetrics and gynecology, and gastrointestinal bleeding. [0004] At present, the clinically used thrombin products are mainly separated and extracted fr...

Claims

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Application Information

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IPC IPC(8): C12N9/74
Inventor 董继胜王金富冯文善魏彦赵波
Owner 黑龙江迪龙制药有限公司
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