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Method for synergetic purification of kallikrein by sodium chloride-ammonia, acetone

A kallikrein and sodium chloride technology, applied in biochemical equipment and methods, peptidase, hydrolase, etc., can solve the problems of tailing, prolonging production cycle, affecting product yield and specific activity, etc. The effect of short cycle, simple operation and low cost

Active Publication Date: 2014-03-26
QINGDAO KANGYUAN PHARMA
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Problems solved by technology

In actual large-scale production, the raw materials contain impurities. If the raw materials or crude products are not pretreated before adopting these high-end chromatographic technologies, the life of the ion exchange medium will be shortened due to the excessive content of impurities in the raw materials, and the Resin adsorption efficiency, and there will be tailing phenomenon in the elution, prolong the production cycle, affect the product yield and specific activity

Method used

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Examples

Experimental program
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Embodiment 1

[0013] After extracting and precipitating porcine pancreas as raw material to make acetone powder, the kinin is prepared by using modern protein purification techniques such as acetic acid extraction, acetone precipitation, ether degreasing and dehydration, sodium chloride-ammonia water, acetone synergistic impurity removal, etc. releasing the enzyme, comprising the steps of:

[0014] 1. Acetic acid extraction: After making 10kg of pig pancreas into acetone powder, add 30 times the amount of acetic acid solution, stir and extract, and centrifuge to obtain 30L of supernatant.

[0015] 2. Precipitation with acetone: collect the supernatant and add acetone for precipitation. The precipitate was defatted with acetone and ether, dehydrated, and dried to obtain 96 g of crude kallikrein.

[0016] 3. Sodium chloride-ammonia water and acetone synergistically remove impurities: dissolve the crude product with 50 times the amount of 8°C sodium chloride solution, adjust the pH to 8.0 wit...

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Abstract

The invention discloses a method for synergetic purification of kallikrein by sodium chloride-ammonia, acetone. A technical scheme provided by the invention employs porcine pancreas as a raw material, which is subjected to extraction and precipitation to obtain an acetone powder, and flexibly uses traditional protein purification technologies such as acetic acid extraction, acetone sediment, diethyl ether degreasing and dehydration, and synergetic purification by sodium chloride-ammonia and acetone to prepare kallikrein. The invention has the characteristics of short production period, simple operation, good chemical and thermal stability, low cost, and is very suitable for mass production.

Description

technical field [0001] The invention relates to the field of biotechnology, specifically extracting porcine pancreas as a raw material, precipitating porcine pancreas to make acetone powder, flexibly using acetic acid extraction, acetone precipitation, ether degreasing and dehydration, sodium chloride-ammonia water, acetone synergistic removal of impurities, etc. Traditional protein purification techniques to prepare kallikrein. Background technique [0002] Kallikrein (kallikein) is a natural enzyme drug isolated and purified from porcine pancreas. It is also called kalikrein clinically. It has the effect of stretching capillaries and arterioles. It is often used clinically to treat hyperlipidemia. Prevention and treatment of fatty liver, atherosclerosis, hypertension, angina pectoris and other diseases, its development has important medical and social value. In my country, there is still a lot of room for development in the production of kallikrein. [0003] In the resea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
CPCC12N9/6445C12Y304/21008
Inventor 刘乃山李静洁刘君张晓涵
Owner QINGDAO KANGYUAN PHARMA
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