Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for purifying urokinase through resin regeneration column efficiency improvement

A urokinase and potency technology, applied in the field of urokinase, can solve the problems of low potency, immature urokinase, low yield, etc., and achieve the effects of improving column efficiency, huge economic and social benefits, and saving costs

Inactive Publication Date: 2017-03-22
QINGDAO KANGYUAN PHARMA
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] But the prior art is immature for the method of purifying urokinase, and its yield is very low, titer is not high, can't be fully applied clinically

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) Rough processing of urokinase

[0035] Weigh 1T of clear urine with a pH of 5.0-6.5, stir constantly, slowly add 16.5kg of chitin to elute with ammonia water after the adsorption is complete, then salt out with 3.5kg of ammonium sulfate, settle overnight, centrifuge and place in distilled water Vacuum dry in a vacuum desiccator using phosphorus as a water-absorbing agent. After drying, the crude urokinase is obtained.

[0036] 2) Adsorption column chromatography (take 0.25L DEAE resin column as an example)

[0037] 2.1) Backwash the DEAE resin in the column with purified water, put it into a bucket, immerse it in purified water and stir well, let it stand for 1 hour, pour off the supernatant and fine particles, repeat three times, and immerse in purified water overnight.

[0038] 2.2) Filter the DEAE resin, wash it with a small amount of purified water, put it into a barrel after filtering it again, immerse it in 0.5 mol / L hydrochloric acid, stir for 1 hour, filter ...

Embodiment 2

[0056] 1) Rough processing of urokinase

[0057]Weigh 1T of clear urine with a pH less than 6.5, stir constantly, slowly add 16.5kg of chitin and elute with ammonia water after the adsorption is complete. Vacuum dry the water-absorbing agent in a vacuum desiccator, and obtain the crude urokinase after drying.

[0058] 2) Adsorption column chromatography (take 0.25L DEAE resin column as an example)

[0059] 2.1) Backwash the DEAE resin in the column with purified water, put it into a bucket, immerse it in purified water and stir well, let it stand for 1 hour, pour off the supernatant and fine particles, repeat three times, and immerse in purified water overnight.

[0060] 2.2) Filter the DEAE resin, wash it with a small amount of purified water, put it into a barrel after filtering it again, immerse it in 0.5 mol / L hydrochloric acid, stir for 1 hour, filter it, wash it with purified water until it is neutral, and then wash it with 0.5 mol / L hydrochloric acid Soak in NaOH solu...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for purifying urokinase through resin regeneration column efficiency improvement. The method particularly comprises the steps that fresh urine of healthy adult females is adopted as a raw material, and the high-purity urokinase can be prepared through the modern protein high-end biochemical separation technologies such as chitin adsorption, ammonium hydroxide elution, ammonium sulfate salting out, adsorption column chromatography and affinity chromatography. The total yield can be increased to 75% or above, the total tier is not lower than 50,000 iu / mg, and the column efficiency is improved by about 55%.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing urokinase with high yield and stable titer by treating and regenerating specific resin to improve column efficiency. Background technique [0002] Urokinase is an enzyme protein isolated from healthy male urine or obtained from human kidney tissue culture. Acting directly on the endogenous fibrinolytic system, it can catalyze the cleavage of plasminogen or plasmin. It can also degrade fibrinogen, coagulation factor V and coagulation factor V3 in blood circulation, thereby exerting thrombolytic effect. It has quick onset and good effect on newly formed thrombus, and can also increase the activity of vascular ADP enzyme, inhibit ADP-induced platelet aggregation, and prevent thrombosis. This product is mainly used for thrombolytic therapy of thromboembolic diseases. Including acute extensive pulmonary embolism, coronary embolism and myocardial infarction within...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/72
CPCC12N9/6456C12Y304/21031
Inventor 刘乃山张晓
Owner QINGDAO KANGYUAN PHARMA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com