Method for preparing high purity chymotrypsin

A high-purity technology of chymotrypsin, applied in the field of biopharmaceuticals, to achieve the effect of simplifying the process, reducing production costs and improving market competitiveness

Inactive Publication Date: 2007-04-11
宁波林叶生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chymotrypsin produced by traditional techniques obviously cannot meet this demand

Method used

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  • Method for preparing high purity chymotrypsin

Examples

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Effect test

Embodiment 1

[0036] Take 150 kg of fresh frozen beef pancreas. These beef pancreas are purchased from professional animal husbandry or meat companies with good quality assurance and complete animal quarantine certificates. The transportation of raw materials is refrigerated and the storage conditions are frozen. Frozen raw materials are crushed directly. Add 450L of pre-cooled 0.25N sulfuric acid solution, transfer to a 1-ton stirring container, stir and extract in a 4°C freezer for 3 hours, and place it overnight. The next day, filter with plate and frame filter equipment instead of bag filter in the traditional process. The separated residue is bagged and treated according to environmental protection requirements. The obtained clear liquid is stirred and added with ammonium sulfate to 30% saturation, and a small amount of diatomaceous earth is added. As a filter aid, place in a 4°C freezer overnight and then filter to obtain the extract. Add ammonium sulfate to the extract to 70% satura...

Embodiment 2

[0038] Take 450 kg of fresh frozen bovine pancreas, and use a pulverizer to directly pulverize the frozen raw materials into a slurry. Add 1350L of pre-cooled 0.25N sulfuric acid solution, transfer to a 2-ton stirring container, stir and extract in a 4°C refrigerator for 5 hours, and place it overnight. The next day, use a plate-and-frame filter to filter. The separated residue is bagged and treated according to environmental protection requirements. Stir and add ammonium sulfate to the clear liquid to 30% saturation, and add a small amount of diatomaceous earth to filter, and place it in a cold storage at 4°C. After overnight, the extract was obtained by filtration. Add ammonium sulfate to the extract to 70% saturation, place it in a 4°C freezer overnight, absorb the supernatant the next day, discard it, and centrifuge to obtain 23 kg of precipitate. Dissolve the precipitate with 34.5 L of ice water, and then add 11.5 kg of saturated ammonium sulfate solution to adjust the p...

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Abstract

The affinity chromatographic process for producing high purity chymotrypsin includes the following technological steps: freezing fresh ox or pig pancreas as the material, crushing and extracting protein, stepped salting out and crystallizing zymogen, enzymolyzing to obtain coarse product, affinity chromatographic separating and purifying, ultrafiltering and concentrating sterilizing, and vacuum freeze drying to obtain product. Compared with available technology, the present invention has the advantages of suitability for large scale produce, high efficiency, specificity, low cost and high product quality.

Description

technical field [0001] The invention relates to a preparation method of chymotrypsin, in particular to a preparation method of producing high-purity chymotrypsin from chymotrypsinogen by using an affinity chromatography process; it belongs to the field of biopharmaceuticals and relates to the biochemical separation of medicinal protease technology. Background technique [0002] Chymotrypsin is a protease isolated and purified from bovine pancreas or porcine pancreas. Its pharmacological and clinical functions mainly include two aspects: 1. Anti-inflammation: it has good curative effect on various inflammations, inflammatory edema, hematoma, ulcer and thrombus. 2. Anti-cancer: It can promote the penetration of anti-cancer drugs into the lesion, and promote the effect of chemotherapy drugs. It is reported that the anti-cancer synergistic effect is more than 50%, and the effect is stronger when it is used locally in large doses. Large-dose use can treat a variety of tumors, s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
Inventor 叶昀林克王洪森
Owner 宁波林叶生物科技有限公司
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