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

Method for preparing human urinary kallidinogenase crude product

A technology for urinary kininogenase and crude products, which is applied in the field of urine protein extraction, can solve problems such as difficult separation and purification, and affect KN purification, and achieve the effects of improving specific activity, low production cost, and reducing difficulty

Active Publication Date: 2013-07-17
YANGZHOU AIDEA BIOTECH
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this method, due to the strong adsorption performance of the adsorbent, more proteins are adsorbed at the same time, which brings greater difficulty to the downstream purification.
Especially KN and UTI, both of which have similar molecular sizes and are both acidic proteins, are difficult to separate by conventional purification methods, which affects the subsequent purification of KN

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 preparing human urinary kallidinogenase crude product
  • Method for preparing human urinary kallidinogenase crude product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 100 kg of macroporous anion exchange resin is used as the adsorbent. After regeneration treatment, the water is flushed to neutral, and put into a filter cloth bag with a pore size of 50 mesh. In the bucket or urinal, make urine or urine diluted with water flow through the resin, collect the resin that absorbs urine protein after 24 hours, wash the adsorbed resin with drinking water, and finally pack it into the column; use 0.1M NaCl was used for washing, 0.8M NaCl was used for elution, and the elution peaks were collected.

[0037] Use an ultrafiltration membrane with a molecular weight cutoff of 30K for ultrafiltration concentration, adjust the pH of the ultrafiltration concentrate to 8.0 and conductance to 2.2mS / cm, and the metal ion that has been balanced on the surface is Cu 2+ The metal chelating affinity chromatography column (Chelating sepharose FF); then use the balance solution (equilibrium solution formula: 0.02M phosphate buffer, 0.2M NaCl, pH8.0) to wash th...

Embodiment 2

[0043] 200 kg of macroporous anion exchange resin is used as the adsorbent. After regeneration treatment, the water is flushed to neutral, and put into a filter bag with a pore size of 50 mesh. In the bucket or urinal, make urine or urine diluted with water flow through the resin, collect the resin that absorbs urine protein after 24 hours, wash the adsorbed resin with drinking water, and finally pack it into the column; use 0.1M NaCl was used for washing; 0.8M NaCl was used for elution, and the elution peaks were collected.

[0044] Concentrate by ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 30K, adjust the pH of the ultrafiltration concentrate to 8.0 and conductance to 2.2mS / cm, and put the balanced Cu 2+ Metal chelate affinity chromatography column; then wash the metal chelate affinity chromatography column with equilibrium solution (equilibrium solution formula: 0.02M phosphate buffer, 0.2M NaCl, pH8.0), collect the sample penetration...

Embodiment 3

[0050] 100 kg of macroporous anion exchange resin is used as the adsorbent. After regeneration treatment, the water is flushed to neutral, and put into a filter cloth bag with a pore size of 50 mesh. In the bucket or urinal, make urine or urine diluted with water flow through the resin, collect the resin that absorbs urine protein after 24 hours, wash the adsorbed resin with drinking water, and finally pack it into the column; use 0.1M NaCl was used for washing, 0.5M NaCl was used for elution, and the elution peaks were collected.

[0051] Use an ultrafiltration membrane with a molecular weight cutoff of 30K for ultrafiltration concentration, adjust the pH of the ultrafiltration concentrate to 6 and the conductivity to 0.5mS / cm, and the metal ion that has been balanced on the surface is Zn 2+ The metal chelate affinity chromatography column; then use the balance solution (balance solution formula: 0.01M phosphate buffer, pH6.0) to wash the metal chelate affinity chromatography...

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 preparing a human urinary kallidinogenase crude product. The method for preparing the human urinary kallidinogenase crude product comprises the following steps of: after adsorbing urokinase protein in urine for a certain time by using an adsorbing agent, collecting the adsorbing agent in which the urokinase protein is adsorbed and centrally eluting; enabling the eluted urokinase protein solution to pass through a metal chelating affinity chromatography column, wherein a human urinary kallidinogenase inhibiting agent is not adsorbed and the human urinary kallidinogenase is combined onto the metal chelating affinity chromatography column, and the human urinary kallidinogenase inhibiting agent is separated from the human urinary kallidinogenase; and collecting an eluting peak to prepare the human urinary kallidinogenase crude product. Due to the adoption of the technical scheme, the human urinary kallidinogenase is efficiently separated from the humanurinary kallidinogenase inhibition agent in the urine, the subsequent purification treatment for the human urinary kallidinogenase and the human urinary kallidinogenase inhibiting agent is facilitated and the purification yield is increased; and the co-generation of two protein crude products is realized, and thus lower production cost is realized.

Description

technical field [0001] The invention relates to a method for extracting urine protein, in particular to a method for preparing a crude product of human urinary kininogenase. Background technique [0002] There are more than 300 kinds of proteins in human urine. At present, urokinase, human urinary trypsin inhibitor and human urinary kininogenase have been developed and extracted. These components have important therapeutic value in clinic. [0003] Urinary Kallidinogenase (hereinafter referred to as KN) is a glycoprotein composed of 238 amino acids isolated from human urine, with an isoelectric point of about 4 and a molecular weight of about 54000D. It belongs to serine protease. It activates the conversion of human plasma kininogen to kinin. At present, it has been developed as a drug for the treatment of acute cerebral infarction. [0004] Urinary trypsin inhibitor (hereinafter referred to as UTI) is a glycoprotein composed of 143 amino acids isolated and purified from ...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/64
Inventor 苗丕渠苏古方池正昌
Owner YANGZHOU AIDEA BIOTECH
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