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Preparation method of Lumbrokinase

A technology of lumbrokinase and production process, which is applied in the field of lumbrokinase preparation technology, can solve the problems of low lumbrokinase yield, low production capacity, and high production cost, and achieve the effects of reducing waste liquid, reducing production cost, and improving production capacity

Inactive Publication Date: 2010-06-30
QD BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to overcome the shortcomings of the existing lumbrokinase production process: the yield of lumbrokinase is low, the preparation cost is high, there is a lot of waste liquid to be treated, and the production capacity is low. Process method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] (1) Cleaning of fresh worms

[0048] Remove the soil layers on the upper part of the earthworms in the basket or on the ground, spread the earthworms into a thin layer, and let the earthworms discharge the dirt for 10 to 18 hours under the light, rinse twice in the tank with tap water to wash away the mud and sticky matter, Pack in plastic barrels and store in a cold storage for later use.

[0049] (2) Refining

[0050] The earthworms were ground into a slurry with a colloid mill, and the earthworms were homogenized with sodium citrate at pH 5.3 added with 10% ethanol at a ratio of (V / V) 1:4.

[0051] (three) slag removal

[0052] Use a decanter centrifuge to remove the coarse residue in the feed liquid to obtain a supernatant.

[0053] (4) Salting out

[0054] Ammonium sulfate was added to the supernatant at a saturation concentration of 30%, stirred and dissolved completely, and left to stand for 1 hour.

[0055] (5) Centrifugal

[0056] Use a vertical high-speed ...

Embodiment 2

[0068] (1) Cleaning of fresh worms

[0069] Remove the soil layers on the upper part of the earthworms in the basket or on the ground, spread the earthworms into a thin layer, and let the earthworms discharge the dirt for 10 to 18 hours under the light, rinse twice in the tank with tap water to wash away the mud and sticky matter, Pack in plastic barrels and store in a cold storage for later use.

[0070] (2) Refining

[0071] The earthworms were ground into a slurry with a colloid mill, and the earthworms were homogenized with pH 5.0 sodium citrate added with 15% ethanol at a ratio of (V / V) 1:3.

[0072] (three) slag removal

[0073] Use a decanter centrifuge to remove the coarse residue in the feed liquid to obtain a supernatant.

[0074] (4) Salting out

[0075] Ammonium sulfate was added to the supernatant at a saturation concentration of 30%, stirred and dissolved completely, and left to stand for 1 hour.

[0076] (5) Centrifugal

[0077] Use a vertical high-speed c...

Embodiment 3

[0089] (1) Cleaning of fresh worms

[0090] Remove the soil layers on the upper part of the earthworms in the basket or on the ground, spread the earthworms into a thin layer, and let the earthworms discharge the dirt for 10 to 18 hours under the light, rinse twice in the tank with tap water to wash away the mud and sticky matter, Pack in plastic barrels and store in a cold storage for later use.

[0091] (2) Refining

[0092] The earthworms were ground into a slurry with a colloid mill, and the earthworms were homogenized with sodium citrate at pH 5.2 added with 20% ethanol at a ratio of (V / V) 1:4.

[0093] (three) slag removal

[0094] Use a decanter centrifuge to remove the coarse residue in the feed liquid to obtain a supernatant.

[0095] (4) Salting out

[0096] Ammonium sulfate was added to the feed solution at a saturation concentration of 30%, and stirred for complete dissolution and left to stand for 1 hour.

[0097] (5) Centrifugal

[0098] Use a vertical high...

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Abstract

The invention discloses a preparation method of Lumbrokinase, comprising the steps of grinding, deslagging, salting-out, centrifuging, ultra-filtrating, column chromatography isolating, ultra-filtrating, degerming, spray-drying and the like. The preparation method of Lumbrokinase provided by the invention is that ethanol is added as adjuvant to remove hybrid protein by physical settlement and ultrafiltration during the homogenate of earthworms, and then a column chromatography purification is performed so that the capacity of the column is reduced, thereby the yield of the Lumbrokinase is increased, the preparation cost is reduced, the waste fluid to be treated is decreased and the production capacity is increased greatly. The preparation method of Lumbrokinase has the characteristics of good quality controllability and stability of the production process.

Description

technical field [0001] The invention belongs to the technical field of biochemical pharmacy, and relates to a preparation method of lumbrokinase. Background technique [0002] The high morbidity, high mortality and high disability of thrombotic diseases seriously threaten the survival of human beings, and thrombolytic drugs are the first choice for the treatment of this disease. At present, there are common side effects that easily cause bleeding in the thrombolytic drugs used clinically; some are expensive and unbearable for ordinary people; some have poor stability and require high storage conditions, which brings a lot of inconvenience to drug users; The kind of drug administration mode is single, only has oral, or only has intravenous or intramuscular injection, and the scope of use medicine is very limited. [0003] Earthworms are collectively referred to as Dilong in traditional Chinese medicine, and can be divided into different species such as Bibreast worm, Chizi A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
Inventor 路兵董金陵李海珍张慧英
Owner QD BIO PHARMA
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