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

Method for extracting and purifying human relaxin-2 from fermentation liquor

A technology of relaxin and fermentation liquid, which is applied in the field of extraction and purification of polypeptide drugs, can solve the problems of complex process steps, long time consumption, and inability to meet large-scale industrial production, and achieve simplified process steps, easy control of process conditions, and shortened extraction and purification time Effect

Inactive Publication Date: 2014-05-14
济南环肽医药科技有限公司
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent document EP0707650 B1 (priority date June 21, 1993, publication date May 21, 2003) discloses a method for preparing human relaxin-2 by gene recombination technology, in which human relaxin is extracted and purified from fermentation broth Su-2 needs to be extracted and purified four times successively using a cation exchange column, a C18 silica gel column, a cation exchange column, and Sephadex G-15. The process steps are complicated and time-consuming; , published on November 20, 2013) discloses a method for preparing human relaxin-2 using Pichia pastoris, wherein the extraction and purification of human relaxin-2 from the fermentation broth requires repeated microporous membrane filtration for several times , and requires three chromatographic column separation and purification operations, which takes a long time
Although the extraction and purification of human relaxin-2 by the above process steps can improve the product yield and the purity of human relaxin-2 in the final product, it cannot meet the needs of large-scale industrial production due to the complexity of the process steps and the time-consuming

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 extracting and purifying human relaxin-2 from fermentation liquor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Extraction and Purification of Human Relaxin-2 from Fermentation Broth

[0021] 1) Take the fermentation broth, centrifuge at 6000rmp for 15 minutes, and collect the supernatant;

[0022] 2) Add 4 times the volume of 70% acetone to the supernatant in step 1), adjust the pH to 5.0 with acetic acid-sodium acetate buffer solution, add urea to make the concentration of urea in the solution 7mol / L, and let stand at 5°C overnight;

[0023] 3) Centrifuge the solution in step 2) at 14000rmp for 30 minutes, pass through a cellulose acetate ultrafiltration membrane with a molecular weight cut-off of 3000, and collect the concentrated solution;

[0024] 4) Put the concentrated solution of step 3) on a SephadexG-50 column, the column length is 22 cm, the elution solvent is 1 mol / L acetic acid containing 0.15 mol / L sodium chloride, the flow rate is 15 mL / hour, collect the eluate, and vacuum low temperature concentrate;

[0025] 5) The concentrated solution of step 4) is...

Embodiment 2

[0028] Example 2 Extraction and Purification of Human Relaxin-2 from Fermentation Broth

[0029] 1) Take the fermentation broth, centrifuge at 6500rmp for 45 minutes, and collect the supernatant;

[0030] 2) Add 4 times the volume of 70% acetone to the supernatant in step 1), adjust the pH to 5.0 with acetic acid-sodium acetate buffer solution, add urea to make the concentration of urea in the solution 5mol / L, and let stand at 7°C overnight;

[0031] 3) Centrifuge the solution in step 2) at 15000rmp for 20 minutes, pass through a cellulose acetate ultrafiltration membrane with a molecular weight cut-off of 3000, and collect the concentrated solution;

[0032] 4) Put the concentrated solution of step 3) on a SephadexG-50 column, the column length is 22 cm, the elution solvent is 1 mol / L acetic acid containing 0.15 mol / L sodium chloride, the flow rate is 15 mL / hour, collect the eluate, and vacuum low temperature concentrate;

[0033] 5) The concentrated solution of step 4) is...

Embodiment 3

[0036] Example 3 Extraction and Purification of Human Relaxin-2 from Fermentation Broth

[0037] 1) Take the fermentation broth, centrifuge at 5500rmp for 45 minutes, and collect the supernatant;

[0038] 2) Add 4 times the volume of 70% acetone to the supernatant in step 1), adjust the pH to 5.0 with acetic acid-sodium acetate buffer solution, add urea to make the concentration of urea in the solution 12mol / L, and let stand at 3°C overnight;

[0039] 3) Centrifuge the solution in step 2) at 15000rmp for 50 minutes, pass through a cellulose acetate ultrafiltration membrane with a molecular weight cut-off of 3000, and collect the concentrated solution;

[0040] 4) Put the concentrated solution of step 3) on a SephadexG-50 column, the column length is 22 cm, the elution solvent is 1 mol / L acetic acid containing 0.15 mol / L sodium chloride, the flow rate is 15 mL / hour, collect the eluate, and vacuum low temperature concentrate;

[0041] 5) The concentrated solution of step 4) i...

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 extracting and purifying human relaxin-2 from fermentation liquor. The method comprises the steps of centrifuging the fermentation liquor and then filtering the fermentation liquor through a cellulose acetate ultrafiltration membrane having the molecular weight cut-off of 3000, orderly loading the filtered fermentation liquor on SephadexG-50 column and Synchropak RP-P C18 reverse-phase HPLC (High Performance Liquid Chromatography) chromatographic column for chromatographic purification, and then carrying out low-temperature rotary evaporation and concentration, and freeze-drying to obtain purified human relaxin-2. The method for extracting and purifying the human relaxin-2 from the fermentation liquor has the advantages that the steps of extracting and purifying the human relaxin-2 are greatly simplified, the extraction and purification time is shortened and the technological conditions are easy to control, and therefore, the method is applicable to large-scale industrial production.

Description

technical field [0001] The invention relates to a method for extracting and purifying polypeptide medicine, in particular to a method for extracting and purifying human relaxin-2 from fermentation broth. Background technique [0002] Human relaxin-2, also known as relaxin, the English name is Relaxin, and the common name of the drug is Serelaxin. It is a polypeptide drug prepared by genetic recombination. Its structure is composed of two peptide chains, the A chain contains 24 amino acids and there are disulfide bonds in the chain, the B chain contains 29 amino acids, and the A chain and the B chain are connected by two disulfide bonds. Its structural formula is as follows: [0003] [0004] Where pGlu is pyroglutamic acid, Tyr is tyrosine, Ser is serine, Asn is asparagine, Cys is cysteine, His is histidine, Val is valine, Gly is glycine, Thr is threonine Amino acid, Lys is lysine, Arg is arginine, Leu is leucine, Phe is phenylalanine, Asp is aspartic acid, Trp is trypto...

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 Applications(China)
IPC IPC(8): C07K14/64C07K1/36C07K1/34C07K1/20
CPCC07K14/64
Inventor 厉保秋房世红袁强
Owner 济南环肽医药科技有限公司
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