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

Method for purifying ulinastatin on basis of ion exchange resin

The technology of ion exchange resin and ulinastatin, which is applied in the field of medicine, can solve the problems of influence on the yield of ulinastatin, complicated purification process, etc., and achieve the effects of increasing molecular weight cut off, simple purification process and improving accuracy.

Inactive Publication Date: 2018-03-23
GUANGDONG TECHPOOL BIO-PHARMA CO LTD
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But for a certain volume of urine, the more complex the purification process, the purity of ulinastatin can be improved to a certain extent, but the yield of ulinastatin is affected

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for purifying ulinastatin on basis of ion exchange resin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1) Take 1 ton of clarified urine, adjust the pH to 6, filter it with gauze, add 8 kg of chitosan-chitin mixture (chitosan: chitin = 3:1) to the filtrate while stirring, adjust the pH to 4, Stir until the adsorption is complete, filter, discard the urine, wash with water to remove residual urine and impurities until clarification; stir and wash chitosan-chitin with ammonia water, and filter to obtain the filtrate; add ammonium sulfate to the filtrate for salting out, stir to dissolve, and static Place, precipitate, and obtain the crude product of ulinastatin after filtering the obtained solid and drying it;

[0042]2) Add sodium acetate-acetic acid buffer solution of pH=5 to the ulinastatin crude product until completely dissolved, continue to add sodium alginate (molecular weight 20,000), stirring, leaving standstill, centrifuging to obtain the lower sediment 1, adding ammonium sulfate buffer solution to the sediment 1 to make pH=6, ultrafiltration to obtain the retenta...

Embodiment 2

[0048] 1) Take 1 ton of clarified urine, adjust the pH to 6.5, filter it with gauze, add 12 kg of chitosan-chitin mixture (chitosan: chitin = 8:1) to the filtrate while stirring, adjust the pH to 5, Stir until the adsorption is complete, filter, discard the urine, wash with water to remove residual urine and impurities until clarification; stir and wash chitosan-chitin with ammonia water, and filter to obtain the filtrate; add ammonium sulfate to the filtrate for salting out, stir to dissolve, and static Place, precipitate, and obtain the crude product of ulinastatin after filtering the obtained solid and drying it;

[0049] 2) Add sodium acetate-acetic acid buffer solution with pH=5.5 to the crude ulinastatin until completely dissolved, and continue to add sodium alginate (molecular weight 50,000), stirring, standing, and centrifugation to obtain the lower sediment 1, adding ammonium sulfate buffer solution to the sediment 1 to make pH=6.5, and ultrafiltration to obtain the r...

Embodiment 3

[0055] 1) Take 1 ton of clarified urine, adjust the pH to 6.5, filter it with gauze, add 12 kg of chitosan-chitin mixture (chitosan: chitin = 5:1) to the filtrate while stirring, adjust the pH to 4, Stir until the adsorption is complete, filter, discard the urine, wash with water to remove residual urine and impurities until clarification; stir and wash chitosan-chitin with ammonia water, and filter to obtain the filtrate; add ammonium sulfate to the filtrate for salting out, stir to dissolve, and static Place, precipitate, and obtain the crude product of ulinastatin after filtering the obtained solid and drying it;

[0056] 2) Add sodium acetate-acetic acid buffer solution with pH=5 to the crude product of ulinastatin until completely dissolved to obtain mixed solution 1;

[0057] Adjust the pH=6 of the mixed solution 1, pass it through the anion exchange resin AmberliteIRA400 at a speed of 120mL / min, and then use NaH with pH=6 2 PO 4 -Na 2 HPO 4 The mixed solution of buf...

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 belongs to the technical field of medicine, and in particular relates to a method for purifying ulinastatin based on an ion exchange resin. In this method, the crude product of ulinastatin is first obtained by adsorption and separation of the mixture of chitosan and chitin, then the crude product is mixed with alginate, followed by ultrafiltration, ion exchange resin purification, ultrafiltration, and finally the obtained ulinastatin after ultrafiltration The retentate was freeze-dried to obtain pure ulinastatin. The method for purifying ulinastatin based on ion exchange resin provided by the application shortens the purification process, and the yield and purity of ulinastatin are high, wherein the yield of ulinastatin reaches 88%; the specific activity reaches 5100IU / mg protein.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for purifying ulinastatin based on anion exchange resin. Background technique [0002] Ulinastatin is an acidic glycoprotein isolated and purified from male urine. According to literature reports, it is composed of 143 amino acids, with sugar chains on the 10th serine and 45th aspartic acid, and the surface of the structure contains a large number of non-polar hydrophobic groups, such as glycine, alanine, valine , leucine, etc. The molecular weight of ulinastatin is 60-70KD determined by HPLC, 40-50KD determined by SDS-PAGE, and the isoelectric point is between 2-3. [0003] Because the two active functional regions in the structure have a wide range of inhibitory enzyme spectrum, they can simultaneously inhibit the activity of various hydrolytic enzymes such as trypsin, phospholipase A2, hyaluronidase, elastase; and, ulinastatin The low-molecular-weight ...

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): C07K14/81C07K1/36C07K1/30C07K1/22C07K1/18
CPCC07K14/8114
Inventor 郑少亮王旭肖益热
Owner GUANGDONG TECHPOOL BIO-PHARMA CO LTD
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