A kind of preparation method of trypsin

A technology of trypsin and catalase, applied in the direction of trypsin, etc., can solve the problems of uncertain inactivation effect, high cost of ultra-high pressure equipment, high production cost, etc., achieve complete inactivation effect, enhance virus safety, and avoid residual Effect

Active Publication Date: 2022-04-08
苏州良辰生物医药科技有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the inactivation effect of high pressure treatment on some viruses is uncertain, and the ultra-high pressure equipment is expensive and the production cost is high, which is basically not advisable in production practice

Method used

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  • A kind of preparation method of trypsin
  • A kind of preparation method of trypsin
  • A kind of preparation method of trypsin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the preparation of trypsin comprises the following steps of carrying out successively:

[0026] (1) After slicing 100g of pancreas and 5g of duodenum, add 20g of anhydrous acetone, stir and mix, then grind with a colloid mill, and autolyse for 4 hours;

[0027] (2) Add 0.5 g of commercially available hydrogen peroxide solution hydrogen peroxide with a mass concentration of 30%, and stir at 2°C for 8 hours;

[0028] (3) Add 100000u unit catalase and stir at 5°C for 2h;

[0029] (4) Put the treatment solution treated in step (3) into a degreasing bucket, add pre-cooled raw acetone with a temperature of 0~5°C and a specific gravity ≤0.800, degrease for 2 hours, and open the valve to release degreasing acetone;

[0030] (5) Add calcium chloride as an activator to the treatment solution after step (4) for activation, stir evenly, put it into an extraction tank, and activate it for 20 hours at a temperature of 5°C;

[0031] (6) Separate and filter the activate...

Embodiment 2

[0037] It is basically the same as Example 1, except that 8 g of commercially available hydrogen peroxide solution with a mass concentration of 30% is added, and the amount of catalase added is 500,000 u. After testing, the prepared trypsin powder contained 1500u / g trypsin and 10000u / g pancrelipase, and no catalase residue.

[0038] Detection method of virus inactivation effect

[0039] After step (1) and before step (2), add 6.5 log 10 TCID50 / 0.1ml titer of the indicator virus EMCV (encephalomyocarditis virus, the amount of virus solution added is 5% of the volume of the solution, about 5ml of virus solution is added to 100g of pancreas).

[0040] The samples before adding hydrogen peroxide and after adding catalase were taken out and inoculated into Vero cell culture respectively to observe the cytopathic effect (CPE). For the effect of the sample before adding hydrogen peroxide on the cytopathic Figure 4 , the effect of the sample after adding catalase reaction on cytopa...

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Abstract

The invention relates to a preparation method of pancreatin, which comprises grinding the pancreas, and then adding hydrogen peroxide to obtain a mixture, wherein the mass percent concentration of the hydrogen peroxide in the mixture is 0.1%-2%, Stir for 1~8h, add catalase to react for 0.5~2h, then degrease, activate, precipitate, and dry to obtain trypsin. The hydrogen peroxide of the present invention can fully contact with the virus in a relatively short period of time so as to achieve the purpose of virus inactivation. In addition, the residual hydrogen peroxide is decomposed into water and oxygen by catalase, so that there is no hydrogen peroxide in the pancreatin. Residues, thus greatly enhancing the virus safety of drugs, so that pancreatin products can meet the stringent requirements of national pharmacopoeias and animal-derived biochemical drug regulations.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of trypsin. Background technique [0002] Pancreatin is a mixed enzyme preparation extracted from the pancreas of mammals such as pigs, cattle, and sheep. The main components are trypsin, pancreatic amylase, and pancreatic lipase, as well as carboxypeptidase, chymotrypsin, and elastase. , kallikrein and ribonuclease. As early as 1938, pancreatic enzyme products have been used for cystic fibrosis and chronic pancreatitis, and most patients need to take it for life. At present, pancreatin is a digestive aid drug recorded in the pharmacopoeias of various countries, and it is clinically used to treat indigestion, loss of appetite, and digestive disorders caused by hepatopancreatic diseases. Since pancreatin contains various active substances, it can be used as raw material medicine to extract required biochemical substances therefrom. In addition, tryp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/94
CPCC12N9/94
Inventor 殷文静
Owner 苏州良辰生物医药科技有限公司
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