Preparation method for chymotrypsin

A technology of chymotrypsin and protein, applied in the field of biopharmaceuticals, can solve the problems of large amount of media used, expensive affinity chromatography media, and unsuitable for large-scale production, and achieve the effect of large market competitiveness and low cost

Active Publication Date: 2013-04-24
QINGDAO JIULONG BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Chinese patent CN1944641A discloses a preparation method of chymotrypsin, which uses an affinity chromatography process to produce high-purity chymotrypsin from prochymotrypsin, but as analyzed in Chinese patent CN1013025B, there are also some shortcomings. The most important one is the affinity chromatography medium Exp

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0041] Example 1

[0042] 1. Crude chymotrypsin production operation:

[0043] (1) Weigh 4 kg of fresh and frozen bovine pancreas, immediately use tweezers and scissors to remove the fat connective tissue, etc., mince, and weigh.

[0044] (2) Place the minced material in a plastic bucket and immerse it in 6L of 0.125mol / L sulfuric acid solution for 22 hours.

[0045] (3) Filter the impregnated material, and then immerse the filter residue with 4L of 0.125mol / L sulfuric acid solution for 1 hour, and discard the filter residue after filtering.

[0046] (4) Combine the two impregnation liquids, measure 11.4L in volume, put in a plastic bucket, add 2.76 kg of solid ammonium sulfate to 40% saturation, and let stand for 12 hours.

[0047] (5) Filter, save the filtrate, measure 10.2L in volume, and discard the solid.

[0048] (6) Add 2.1 kg of solid ammonium sulfate to the filtrate to reach 70% saturation and leave it for 12 hours.

[0049] (7) Filter, save the solid, weigh 116g, discard the filt...

Example Embodiment

[0074] Example 2

[0075] 1. Crude chymotrypsin production operation:

[0076] (1) Weigh 4 kg of fresh and frozen bovine pancreas, immediately use tweezers and scissors to remove the fat connective tissue, etc., mince, and weigh.

[0077] (2) The minced material is placed in a plastic bucket and immersed in 10L of 0.125mol / L sulfuric acid solution for 28 hours.

[0078] (3) Filter the impregnated material, and then immerse the filter residue with 8L of 0.125mol / L sulfuric acid solution for 2.5 hours, and discard the filter residue after filtering dry.

[0079] (4) Combine the two impregnation liquids, measure the volume of 11.5L, put it in a plastic bucket, add 2.78kg of solid ammonium sulfate to 40% saturation, and let it stand for 15 hours.

[0080] (5) Filter, save the filtrate, measure 10.3L in volume, and discard the solid.

[0081] (6) Add 2.1 kg of solid ammonium sulfate to the filtrate to reach 70% saturation, and leave it for 15 hours.

[0082] (7) Filter, save the solid, weigh 11...

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Abstract

The invention provides a preparation method for chymotrypsin. The preparation method comprises the following process procedures: (1), grinding raw material and extracting protein; (2) salting out in a grading manner to obtain a crude product of chymotrypsin; (3), feeding the crude product and crystallizing in a multiple manner; (4), activating; (5), salting out; (6), dialyzing; and (7), freeze-drying. According to the invention, a set of large-scale manufacturing technology is established and has the advantages of simplicity, low cost and the like; the titer of a fine product of prepared chymotrypsin is more than 1,200 unit/milligram; and the prepared chymotrypsin meets the requirements of Chinese pharmacopeia (2010) and has very great market competitiveness.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and designs a biochemical separation technology of medicinal chymotrypsin. Specifically, techniques such as salting out, multiple crystallization combined with dialysis are used to extract chymotrypsin. 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, promote the effect of chemotherapy drugs, and the effect is stronger when used locally in large doses. Large-dose use can treat a variety of tumors, such as breast cancer, cervical cancer, gastric cancer, etc., without any toxic side effects or adverse reactions. [0003] Chinese patent CN1944641A ...

Claims

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Application Information

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IPC IPC(8): C12N9/76
Inventor 朱琳葛翠凤刘翠珍吕金花
Owner QINGDAO JIULONG BIO PHARMA
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