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Method for removing endotoxin in porcin thrombin by gel molecular sieve

A pig thrombin, thrombin technology, applied in biochemical equipment and methods, enzymes, enzymes and other directions, can solve the problems of poor stability and low purity of thrombin

Inactive Publication Date: 2012-06-13
JILIN UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0004] At present, the thrombin used clinically is mostly limited to its normal use due to problems such as low purity, pyrogen content, and poor stability of thrombin. The above problems are also scientific problems that need to be solved urgently. In addition, research and improve pig blood extraction. Stable and high-purity thrombin technology can not only protect the environment, turn waste into wealth and create benefits, but also increase the added value of pig blood, promote employment, and stimulate local economic development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0013] 1 Separation of plasma

[0014] Add 3.0% trisodium citrate to anticoagulant fresh pig blood qualified for quarantine, and stir slowly to reduce blood cell fragmentation. Blood was separated using a high-speed refrigerated centrifuge, centrifuged at 4500 r / min for 10 min at 4°C, and the plasma was collected and stored at 4°C for future use.

[0015] 2 Fibrinogen removal

[0016] The collected plasma was frozen at -20°C for 24 hours, taken out and thawed at 4°C, at this time the fibrinogen precipitated into flocculent precipitates, the precipitates were removed, and the liquid was collected. The total activity of the resulting thrombin was 3.7% higher than that of unfrozen.

[0017] 3 Preparation and activation of thrombin

[0018] Add 1mol / L barium chloride solution to the liquid collected in the second step, the addition amount is 8%, stir for 20min, collect the barium citrate precipitate by suction filtration, dissolve the precipitate in pH8.00.2mol / L EDTA solution ...

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PUM

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Abstract

The invention discloses a method for removing endotoxin in porcin thrombin by a gel molecular sieve, which belongs to the field of deep processing of agricultural and sideline products. The method includes steps of centrifugally separation, column chromatography, enzymatic activation and separation by the molecular sieve so as to remove the endotoxin. The method has the advantages of fine separation effect and high safety in clinical use, and thrombin is high in activity, heat source substances and endotoxin in the thrombin are few.

Description

technical field [0001] The invention belongs to the field of deep processing of agricultural and sideline products, and is an exploratory application research of biotechnology in the deep processing of agricultural and sideline products. Background technique [0002] Thrombin is a natural component in the body's blood coagulation system. It is derived from animal plasma. Thrombin is widely used clinically. It is often applied locally as dry powder or solution to wounds and surgical sites to control capillary blood leakage and is mostly used for bone bleeding. , tonsillectomy and tooth extraction bleeding, etc., and sometimes it can also be taken orally for gastric and duodenal bleeding. Thrombin has a good local hemostatic effect and has no side effects. [0003] According to the global survey conducted by the World Federation of Hemophilia in 2008, based on the consumption of 0.08 units of coagulation factor per capita in countries with the lowest income per capita, my cou...

Claims

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Application Information

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IPC IPC(8): C12N9/74
Inventor 潘风光方珍于挺敏姚刚吴涵王楠
Owner JILIN UNIV
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