Method for separating and extracting alfapsin

Abstract

The invention discloses a method for separating and extracting chymotrypsin, comprising the following steps of: pulverizing and filtering beef pancreas gland and extracting a filtrate; removing hetero-protein from the filtrate, and refining and concentrating the filtrate; adding ammonium sulphate and regulating pH value with sodium hydroxide for culture to separate chymotrypsinogen by crystallization; carrying out extraction and filtration to the culture to extract chymotrypsinogen; recycling the extracting and the filtering liquid of the culture for later use; adding trypsinase to activate zymohydrolysis; and obtaining a first semi-finished product solution after filtering; filling chromatographic column with CM-sopharose FF gel medium; balancing the chromatographic column with acetic acid buffer solution; then loading the extracting and filtering liquid of the culture after balancing; washing the column with balancing liquid until effluent liquid OD(280) value is less than 0.1; eluting the column with monosodium orthophosphate -disodium hydrogen phosphate buffer liquid; and collecting the eluting peak to obtain a solution which is a second semi-finished product; mixing the first semi-finished product with the second semi-finished product; subjecting the mixture to desalting and concentration, aseptic filtration and freeze vacuum drying; and obtaining the finished product of chymotrypsin. The method solves the problems of expensive affinity chromatography medium and less times of repeated use, and is applicable for large scale production.

Description

technical field [0001] The invention relates to a separation and extraction method of chymotrypsin, belongs to the field of biopharmaceuticals, and relates to a biochemical separation technology of medicinal protease. 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 is effective for various inflammations, inflammatory edema, hematoma, ulcers and thrombus. Better curative effect. 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, such as breast cancer, cervical cancer, gastric cancer, etc., without any toxic side effects or adverse reactions. [00...

AI Technical Summary

Problems solved by technology

[0004] In view of the problems existing in the prior art, the purpose of the present invention is to provide a method for separating and extracting chymotrypsin, which can solve the disadvantages of expensive affinity chromatography medium and few reusable times; it can reduce the burden of chromatography procedures, reduce The amount of medium used not only reduces investment and production costs, but also can obtain high specific activity of chymotrypsin, which is suitable for large-scale production

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