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Method for extracting hirudin by stimulating living leech with electricity

An electrical stimulation, hirudin technology, applied in the direction of leech inhibitors, protease inhibitors, specific peptides, etc., can solve problems such as unavoidable seasonal problems of hirudin, excessive dependence on leech resource supply, and unsuccessful realization.

Inactive Publication Date: 2013-03-06
南宁乙翔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the methods for industrial production of hirudin are to obtain hirudin by destroying the living leech, which affects the purity of the obtained hirudin. At the same time, it is too dependent on the supply of leech resources, and it is especially difficult to avoid the seasonal problem of hirudin production.
Utilize the stimulating effect of electric current on the glands of organisms, directly use DC weak current to stimulate living leeches to extract high-purity hirudin in batches and maintain the living traits of leeches in order to simplify the production process of hirudin, improve product purity, and maintain wild leeches The method of reusing resources, reducing production costs, and overcoming seasonal obstacles in production has not yet been successfully realized.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] At room temperature of 20-30°C, first take the live leech from the feeding tank to the storage tank, rinse with deionized water for 2 minutes, remove the impurities on the surface of the living leech, filter out the water, and then immerse it in physiological saline for 20 seconds to make the leech The surface is covered with electrolyte, filtered to remove salt water, and placed in a 200-mesh rectangular stainless steel filter tank. The filter tank is insulated from the ground. Under the filter screen is a sterilized and clean utensil of the same area. To the minimum output current, then turn on the power switch, adjust the current to 1 ~ 6mA, after 40 minutes, disconnect the power supply, collect the liquid dripping from the filter tank in the vessel, and then put the leech from the filter tank back into the storage tank, and then put back to the feeding tank.

[0020] Most of the liquid dripping from the filter tank in the container is the active ingredient of hirudi...

Embodiment 2

[0022] At room temperature of 20-30°C, first take the living leech from the feeding tank to the storage tank, rinse with deionized water for 2 minutes to remove the impurities on the surface of the living leech, filter out the water, and then rinse with normal saline for 20 seconds to make the surface of the leech Cover the electrolyte, filter out the salt water, and place it in a rectangular stainless steel filter tank with a 200-mesh filter at the bottom. The filter tank is insulated from the ground. The same area of ​​sterilized and clean utensils are under the filter screen. The two ends of the filter tank are connected to the two poles of the direct current , first adjust to the minimum output current, turn on the power switch, adjust the current to 5 ~ 12mA, after 20 minutes of electrical stimulation, disconnect the power supply, collect the liquid dripping from the filter tank in the vessel, and then put the leech back from the filter tank storage tank, and then put back...

Embodiment 3

[0025] At room temperature of 20-30°C, first take the live leech from the feeding tank to the storage tank, rinse with deionized water for 2 minutes to remove the impurities on the surface of the live leech, filter out the water, and then immerse it in physiological saline for 20 seconds to cover the surface of the leech. Electrolyte, filter out salt water, put it flat into a rectangular stainless steel filter tank with a 200-mesh filter screen at the bottom, the filter tank is insulated from the ground, and under the filter screen is a sterilized clean vessel of the same area. First adjust to the minimum output current, turn on the power switch, adjust the current to 12-20mA, after 10 minutes of electrical stimulation, disconnect the power supply, collect the liquid dripping from the filter tank in the vessel, and then put the leech from the filter tank for storage tank and put it back into the rearing tank.

[0026] Most of the liquid dripping from the filter tank in the ves...

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PUM

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Abstract

The invention relates to a method for extracting hirudin by stimulating a living leech with electricity. By utilizing the simulating effect of current on organism glands, a way for simulating a living leech to extract high-purity hirudin at mass by direct weak current is directly used, and the living character of the leech is maintained, therefore, the production process of hirudin is simplified, the product purity is improved, the wild leech resources are recycled, the production cost is reduced and the problem of seasonal disorder of production is solved. The hirudin extracted by using the method is subjected to nanofiltration water removal and decompression spray drying to be made into powdery hirudin with the antithrombin activity of 800AT-U / g and liquid hirudin with the antithrombin activity of 1,000AT-U / g when protective agents are added in the liquid hirudin.

Description

technical field [0001] The invention relates to a method for extracting hirudin by stimulating the living leech with electricity. The leech glands release glandular fluid by stimulating the living leech with weak electricity, and at the same time, the leech is ensured to maintain a living state, so as to obtain high levels of Method for the purity of hirudin. Background technique [0002] Hirudin is named after it was first isolated from leeches. Natural hirudin consists of 65 amino acid residues and has a molecular weight of 7000 Daltons. Hirudin can irreversibly combine with the catalytic center of thrombin and the binding center of fibrinogen to inactivate thrombin. It is the most powerful thrombin inhibitor found so far. In addition, it has a strong antithrombotic effect and has inhibitory effect on a variety of experimental thrombus models. Hirudin has preventive and therapeutic effects on various thrombotic diseases such as venous thrombosis, disseminated intravascu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/815C07K1/14
Inventor 苏国强刁建中
Owner 南宁乙翔生物科技有限公司