Separation method of hirudin

A separation method and technology of hirudin, applied in the preparation method of peptides, from leech inhibitors, chemical instruments and methods, etc., can solve the problems of cumbersome extraction steps, low utilization rate of leeches, low activity of hirudin, etc., and achieve environment-free Pollution, improved utilization, high activity effect

Inactive Publication Date: 2017-07-07
佛山市爱的生物科技有限公司
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Problems solved by technology

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a method for separating hirudin, which aims to solve the problems of cumbersome extraction steps, low leech utilization rate, and low activity of hirudin in the existing hirudin extraction method.

Method used

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  • Separation method of hirudin
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Examples

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Effect test

Embodiment 1

[0034] Take 180ml of leech saliva (or leech gastric juice), add 200ml of normal saline, and stir in a 50°C water bath for 10 minutes; transfer the sample to a centrifuge tube with a filter membrane for centrifugation. min; stand still for 20 min, take the supernatant and freeze-dry (-40°C, 35 h) to obtain 10 g of hirudin freeze-dried powder.

Embodiment 2

[0036] Take 440ml of leech saliva (or leech gastric juice), add 2000ml of normal saline, and stir in a 37°C water bath for 10 minutes; transfer the sample to a centrifuge tube with a filter membrane for centrifugation. min; stand still for 20 min, take the supernatant and freeze-dry (-40°C, 20 h) to obtain 23 g of hirudin freeze-dried powder.

Embodiment 3

[0038] Take 500ml of leech saliva (or leech gastric juice), add 250ml of normal saline, and stir in a 40°C water bath for 10 minutes; transfer the sample to a centrifuge tube with a filter membrane for centrifugation. min; stand still for 20 min, take the supernatant and freeze-dry (-40°C, 10 h) to obtain 25 g of hirudin freeze-dried powder.

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Abstract

The invention discloses a separation method of hirudin, which comprises the following steps of adding saline water in saliva or gastric juice of hirudo for diluting; heating in water bath at the temperature of 37 DEG C to 100 DEG C for 3 to 300min, and heating a sample to 36 to 38 DEG C; transferring the sample to a centrifuge tube with a filter membrane for carrying out centrifugal treatment, wherein the molecular weight cutoff of the filter membrane is 5000 to 10000kDa; standing filter liquor obtained through centrifugation, and getting supernate; freeze drying the supernate to obtain hirudin freeze-dried powder. The method provided by the invention is simple in steps, high in efficiency and high in activity of the hirudin, and can reach to 400U / g or above; meanwhile, a toxic solvent has no need to be adopted at all, so that the medicine purity is ensured, the pollution to the environment is avoided, and the separation method is suitable for industrialized production.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, and mainly relates to a method for separating hirudin. Background technique [0002] Thrombus is the main cause of cardiovascular and cerebrovascular diseases, and thrombin is the main cause of thrombus formation. Hirudin is the strongest specific inhibitor of thrombin found so far, which can specifically bind with thrombin to make it lose its coagulation activity. At present, hirudin is mainly extracted from Japanese medical leeches, Philippine leeches, European medical leeches, Indian leeches, and Amazon giant leeches. Clinical experiments have shown that hirudin can not only melt thrombus to eliminate or alleviate the condition, but also prevent thrombus formation, and has an excellent effect of preventing and treating thrombotic cardiovascular disease. [0003] At present, there are many methods for extracting hirudin at home and abroad. For example, in 1970, Markwardt used ethanol fracti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/815C07K1/14
CPCC07K14/815
Inventor 汤浩强
Owner 佛山市爱的生物科技有限公司
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