Purification method of recombinant human vascular endothelial growth factor receptor-antibody fusion protein

A technology of growth factor receptor and vascular endothelium, which is applied in the field of purification of recombinant human vascular endothelial growth factor receptor-antibody fusion protein, which can solve the problems of complex structure and difficult purification work

Active Publication Date: 2015-12-23
QILU PHARMA CO LTD
View PDF4 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Recombinant human vascular endothelial growth factor receptor-antibody fusion protein is a fusion protein with a relatively complex structure. It has many disulfide bonds and glycosylation sites, and contains many basic charge isomers, which brings difficulties to downstream purification work. come very difficult

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
  • Purification method of recombinant human vascular endothelial growth factor receptor-antibody fusion protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A method for purifying recombinant human vascular endothelial growth factor receptor-antibody fusion protein, the specific steps of which are:

[0054] Affinity chromatography

[0055] (1) Equilibrate the rProteinAFastFlow affinity chromatography column (5.0×20cm) with 1000mL disodium hydrogen phosphate-sodium dihydrogen phosphate equilibrium buffer containing 1mol / L NaCl at a concentration of 10mmol / L and a pH value of 7.5, and the recombinant The cell culture supernatant of human vascular endothelial growth factor receptor-antibody fusion protein was loaded as a sample, so that the recombinant human vascular endothelial growth factor receptor-antibody fusion protein was adsorbed on the affinity chromatography medium, and after loading, continue to use 1mol / LNaCl concentration of 10mmol / L, pH value of 7.5 disodium hydrogen phosphate-sodium dihydrogen phosphate equilibrium buffer for washing, and then citric acid-sodium citrate buffer solution with a concentration of 2...

Embodiment 2

[0070] A method for purifying recombinant human vascular endothelial growth factor receptor-antibody fusion protein, the specific steps of which are:

[0071] Affinity chromatography

[0072] (1) Equilibrate the Mabselect affinity chromatography column (5.0×20cm) with 1000mL disodium hydrogen phosphate-sodium dihydrogen phosphate equilibration buffer containing 1mol / LNaCl at a concentration of 10mmol / L and a pH value of 7.0, and the recombinant The cell culture supernatant of human vascular endothelial growth factor receptor-antibody fusion protein was loaded as a sample, so that the recombinant human vascular endothelial growth factor receptor-antibody fusion protein was adsorbed on the affinity chromatography medium, and after loading, continue to use 1mol / LNaCl concentration of 10mmol / L, pH value of 7.0 disodium hydrogen phosphate-sodium dihydrogen phosphate equilibrium buffer for washing, and then use concentration of 10mmol / L, pH value of 5.5 citric acid-sodium citrate b...

Embodiment 3

[0087] A method for purifying recombinant human vascular endothelial growth factor receptor-antibody fusion protein, the specific steps of which are:

[0088] Affinity chromatography

[0089] (1) MabselectSure affinity chromatography column (5.0 × 20cm) is equilibrated with 1000mL disodium hydrogen phosphate-sodium dihydrogen phosphate equilibrium buffer solution containing 0.9mol / LNaCl concentration of 20mmol / L and pH value of 7.5, and The cell culture supernatant of the recombinant human vascular endothelial growth factor receptor-antibody fusion protein was loaded as a sample, so that the recombinant human vascular endothelial growth factor receptor-antibody fusion protein was adsorbed on the affinity chromatography medium. The concentration of 0.9mol / LNaCl is 20mmol / L, the pH value is 7.5 disodium hydrogen phosphate-sodium dihydrogen phosphate equilibrium buffer solution for washing, and then the concentration is 30mmol / L, the pH value is 6.0 citric acid-sodium citrate Bu...

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 relates to production of a genetic engineering drug by a recombinant DNA technology, belongs to the biotechnical field, and particularly relates to a purification method of a recombinant human vascular endothelial growth factor receptor-antibody fusion protein, that is to say, a cell supernatant containing the recombinant human vascular endothelial growth factor receptor-antibody fusion protein is successively subjected to affinity chromatography, anion exchange chromatography, cation exchange chromatography, hydrophobic chromatography and desalination chromatography to obtain the high-quality recombinant human vascular endothelial growth factor receptor-antibody fusion protein. The purification method has the characteristics of low cost, high purity of the target protein, easily controlled technological process, easily large-scale enlarged production and the like.

Description

technical field [0001] The invention relates to the production of genetic engineering drugs by using recombinant DNA technology, in particular to a purification method of recombinant human vascular endothelial growth factor receptor-antibody fusion protein, which belongs to the field of biotechnology. Background technique [0002] Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) stimulates the formation of new blood vessels by combining with a specific receptor - vascular endothelial growth factor receptor (vascular endothelial growth factor receptor, VEGFR). In the process of tumor growth, the number of VEGFs surges, and the regulation imbalance between VEGFs and angiogenesis inhibitors greatly promotes the division, proliferation and migration of endothelial cells, improves vascular permeability, and provides a good microenvironment for tumor growth and metastasis. . [0003] Recombinant human vascular endothelial growth factor receptor-antibo...

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): C07K19/00C07K1/22C07K1/20C07K1/18C07K1/16
Inventor 孙丽霞陈俊良孙波邢婷婷贾国栋朱鹏飞王庆民郑焕兰丁仁志石干
Owner QILU PHARMA CO LTD
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