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

A Mixed Mode Based Expanded Bed Adsorption Method for Separation of Human Serum Albumin

A technology of human albumin and mixed mode, which is applied in the field of expanded bed adsorption separation of human albumin based on mixed mode, can solve the problems of decreased concentration of target substances, prolonged process time, and reduced adsorption capacity, so as to reduce degradation, Simplified preprocessing steps, easy to scale up effects

Active Publication Date: 2020-04-10
ZHEJIANG UNIV
View PDF11 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cation exchange method is adopted in the protein capture stage, and the feed liquid needs to be adjusted to pH 4.0 and diluted. The acidic environment is easy to activate protease, resulting in the degradation of human serum albumin. Dilution will reduce the concentration of the target substance, reduce the adsorption capacity, and increase the processing capacity. , the process time is prolonged

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
  • A Mixed Mode Based Expanded Bed Adsorption Method for Separation of Human Serum Albumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Take the Pichia fermented liquid containing recombinant human serum albumin, adjust the pH to 6.0, add sodium octanoate to 5 mM, and heat at 68° C. for 30 minutes to obtain a crude human serum albumin solution. The mixed-mode adsorbent (agarose microspheres with quartz sand as a weighting agent, L-tryptophan as a ligand) is packed into an expanded bed with an inner diameter of 1 cm, a column height of about 20 cm, and pH 5.0 acetic acid-sodium acetate buffer Pre-equilibrate to reach 2.0 times the expansion rate, keep for 20 minutes; adjust the pH of the crude human serum albumin solution to 5.0, and load 10ml of the sample. Wash 5 times column volume with equilibration buffer, wash 10 times column volume with 20mM disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (pH 7.5), wash 10 times column volume with 20mM acetic acid-sodium acetate buffer (pH 4.0), The eluted II fraction was collected, desalted, and freeze-dried to obtain human serum albumin. The purit...

Embodiment 2

[0025]Take the Pichia fermented liquid containing recombinant human serum albumin, adjust the pH to 6.0, add sodium caprylate to 10 mM, and heat at 68° C. for 30 min to obtain a crude human serum albumin solution. Mixed mode adsorbent (agarose microspheres with titanium dioxide added as a weighting agent, L-tryptophan methyl ester as a ligand) is packed into an expanded bed with an inner diameter of 1 cm, a column height of about 20 cm, pH 5.5 acetic acid-sodium acetate The buffer was pre-equilibrated to achieve a 1.8-fold expansion rate, and kept for 20 minutes; the pH of the crude human serum albumin solution was adjusted to 5.5, and 10ml of the sample was loaded. Wash 5 times the column volume with equilibration buffer, wash 5 times the column volume with 20mM disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (pH 7.0), wash with 10 times the column volume with 20mM acetic acid-sodium acetate buffer (pH 3.5), The eluted II fraction was collected, desalted, and f...

Embodiment 3

[0027] Take the Hansenula fermented liquid containing recombinant human serum albumin, adjust the pH to 6.0, add sodium octanoate to 15 mM, and heat at 68° C. for 30 min to obtain a crude human serum albumin solution. Mixed mode adsorbent (agarose microspheres added with tungsten carbide as a weighting agent, L-tryptophan ethyl ester as a ligand) is packed into an expanded bed with an inner diameter of 1 cm, the column height is about 20 cm, pH 6.0 disodium hydrogen phosphate - Sodium dihydrogen phosphate buffer pre-equilibrated to achieve 2.2 times the expansion rate, keep for 20 minutes; adjust the pH of the crude human serum albumin solution to 6.0, and load 10ml of the sample. Wash 5 times column volume with equilibration buffer, wash 15 times column volume with 20mM disodium hydrogen phosphate-citrate buffer (pH8.0), and elute with 10 times column volume with 20mM citric acid-sodium citrate buffer (pH 4.5) , collected the eluted II fraction, desalted, and freeze-dried to ...

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

PropertyMeasurementUnit
purityaaaaaaaaaa
purityaaaaaaaaaa
clearance rateaaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for adsorbing and separating human serum albumin by expanded bed adsorption based on a mixed mode, which can directly separate and recombine human serum albumin from a yeast fermentation solution, and belongs to the protein separation technology in the biochemical field. The method comprises the following steps: (1) preprocessing a fermentation solution; obtaining the yeast fermentation solution containing the human serum albumin, adding sodium caprylate, and heating and deactivating protease; (2) performing expanded bed adsorption, directly separating the yeast fermentation solution by using an expanded bed filled with a mixed-mode adsorption agent, and collecting elution peaks; and (3) desalting and drying: desalting a collected solution, freeze drying to obtain the human serum albumin with the purity greater than 95 percent. The invention is characterized by developing a novel separation process, so that the high-purity recombined human serum albumin can be directly separated from the yeast fermentation solution. A critical point of the invention lies in adopting the expanded bed adsorption agent based on the mixed mode, cells do not need to be removed from the yeast fermentation solution, the ion strength does not need to be adjusted, the elution condition is moderate, and the method has the characteristics of simple process, high separation efficiency and high yield.

Description

technical field [0001] The invention belongs to the technical field of protein separation, and relates to a mixed mode-based expanded bed adsorption separation method for human serum albumin. Background technique [0002] Human albumin is the most abundant protein in human plasma, accounting for about 60% of plasma protein. Its main physiological function is to maintain plasma osmotic pressure, combine and transport nutrients. It is clinically used in the treatment of hemorrhagic shock, traumatic shock, acute hypovolemia and hypoalbuminemia. It can also be used as an additive for cell culture, drug adjuvant, excipient, etc., and has a wide range of application values. [0003] Clinically used human serum albumin products are mainly extracted and purified from human plasma. The preparation methods include cold ethanol precipitation, ammonium sulfate precipitation, rivanol precipitation, caprylate precipitation, etc. The process is complicated, and viruses or other potential ...

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): C07K14/765C07K1/14
CPCC07K14/765
Inventor 林东强吴启赐姚善泾
Owner ZHEJIANG UNIV
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