Method for purifying Fc fusion protein

A fusion protein, purification method technology, applied in chemical instruments and methods, fusion polypeptides, animal/human proteins, etc., can solve the problems of easy formation of degradation and multimerization, poor stability of Fc fusion proteins, etc.

Inactive Publication Date: 2019-08-27
北京军科华仞生物工程技术研究有限公司
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Fc fusion proteins are less stable in animal cell expression supernatants and are prone to degradation and multimerization

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 Fc fusion protein
  • Method for purifying Fc fusion protein
  • Method for purifying Fc fusion protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] A purification method for purifying an Fc fusion protein according to this embodiment specifically includes the following steps:

[0041] 1. First capture the GLP-1-Fc fusion protein with Mabselect SuRe LX:

[0042] 1.1. Balance 4 column volumes of the chromatography column with affinity balance solution (25mM Tris, 25mM NaCl, pH7.5);

[0043] 1.2. Sample loading, sample loading retention time is 6min;

[0044] 1.3. After loading the sample, rebalance 3 column volumes with affinity balance solution, and then wash 4 column volumes with eluent (25mM Tris, 0.3M NaCl, pH7.5);

[0045] 1.4. Then wash the chromatography column with 50mM acetate, pH4.5 for 3 column volumes, elute the sample with 50mM acetate, pH=3.5, collect the main peak at 280nm at ultraviolet, and collect the peak parameters: start collecting as UV =80mAU, terminated UV collection=80mAU;

[0046] 1.5. Wash 3 column volumes with 0.1M NaOH, equilibrate with a balance solution until the pH is stable, and st...

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 Fc fusion protein. The method is characterized by comprising the following steps: (1) capturing GLP-1-Fc fusion protein by using Mabselect SuRe LX; (2) further removing degradation fragments by using Capto Q ImpRes ion exchange chromatography; (3) removing a large quantity of aggregates with CHT I type filler. The method disclosed by the invention iscapable of removing the degradation and aggregates in interest protein.

Description

technical field [0001] The present invention relates to an Fc fusion protein, more specifically, it relates to a purification method for purifying the Fc fusion protein. Background technique [0002] Fc fusion protein refers to a new type of protein produced by fusing a certain biologically active functional protein molecule with the Fc fragment of IgG by using genetic engineering and other technologies. After the functional protein molecule is connected to the Fc fragment, the stability of the functional protein is improved, and the half-life in vivo is increased. The Fc fragment can be fused with one or two different functional fragments to form highly effective targeted drugs. This type of protein not only retains the biological activity of the functional protein, but also retains the ADCC and CDC effects mediated by the Fc fragment. Among the top 15 global drug sales in the first half of 2018, sales of Fc fusion protein drugs (Enbrel of Amgen / Pfizer and Eylea of ​​Bayer...

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): C07K19/00C07K1/22C07K1/18C07K1/16
CPCC07K14/605C07K2319/30
Inventor 游文龙李文宾
Owner 北京军科华仞生物工程技术研究有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap