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

Method for purifying antibody through cation exchange chromatography

A technology of cation exchange and chromatography, which is applied in the field of antibody purification, can solve the problems of low packing capacity, long operation time, and low packing utilization rate, and achieve low packing capacity, long operating time, and low packing utilization rate. Effect

Inactive Publication Date: 2019-02-15
HJB HANGZHOU CO LTD
View PDF6 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a method for purifying antibodies by cation exchange chromatography, to solve the technical problems of low filler loading, long operation time and low filler utilization in the prior art. The purification method passes through the cation exchange layer The analytical method improves the removal ability of HCP and polymers, saves process operation time and reduces costs

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 antibody through cation exchange chromatography
  • Method for purifying antibody through cation exchange chromatography
  • Method for purifying antibody through cation exchange chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A cation chromatography membrane Mustang S XT Acrodis Units (PALL) was used with a membrane volume of 0.86 mL and an operating flow rate of 8.6 mL / min. The source of the sample is the monoclonal antibody (Mab1) expressed by CHO cells, which is captured by protein A affinity chromatography. After low pH inactivation, the pH of the sample antibody solution is adjusted to 5.5, and the conductivity value is 5mS / cm. The antibody solution is obtained after deep filtration in the middle . In the obtained antibody solution, the Mab1 concentration was 10.4 g / L, the purity was 97.0%, the multimer content was 2.5%, and the HCP content was 212.7 ppm.

[0042] After the chromatographic membrane was equilibrated with equilibration buffer (50mM NaAc-HAc, pH 5.5), 7.2mL diluted sample Mab1 concentration of 1.8g / L, purity of 97.0%, multimer content of 2.5% antibody solution was loaded. 15g / L MV. Use elution buffer (50mM NaAc-HAc, 0.5M NaCl, pH 5.5) for gradient elution (0-100%, 30MV),...

Embodiment 2

[0054] A cation chromatography membrane Mustang S XT Acrodis Units (PALL) was used with a membrane volume of 0.86 mL and an operating flow rate of 8.6 mL / min. The source of the sample is the monoclonal antibody (Mab1) expressed by CHO cells, which is captured by protein A affinity chromatography. After low pH inactivation, the pH of the sample antibody solution is adjusted to 5.5, and the conductivity value is 5mS / cm. The antibody solution is obtained after deep filtration in the middle . In the obtained antibody solution, the Mab1 concentration was 4.1 g / L, the purity was 97.0%, the multimer content was 2.5%, and the HCP content was 212.7 ppm.

[0055] Use equilibration buffer (50mM NaAc-HAc, pH 5.5, conductivity value 3.2mS / cm) to equilibrate the chromatographic membrane for 30MV. After loading 5.2mL of sample, the sample begins to break through and collect the flow-through. Continue to load the sample until 279mL is loaded, that is, when the antibody load is 1328g / LMV, the...

Embodiment 3

[0061] Prepacked column Fractogel COO-(Millipore), diameter 0.8cm, column height 10cm, column volume 5mL, operating linear flow rate 300cm / h. The source of the sample is the monoclonal antibody (Mab1) expressed by CHO cells, which is captured by protein A affinity chromatography. After low pH inactivation, the pH of the sample antibody solution is adjusted to 5.5, and the conductivity value is 5mS / cm. The antibody solution is obtained after deep filtration in the middle . In the obtained antibody solution, the Mab1 concentration was 10.4 g / L, the purity was 97.0%, the multimer content was 2.5%, and the HCP content was 212.7 ppm.

[0062] Equilibrate the chromatography column with equilibration buffer (50mM NaAc-HAc, pH 5.5) for 3CV, and after loading 32mL of the sample, the sample begins to break through, and the flow-through solution is collected. Continue to load the sample until 495mL is loaded, that is, when the antibody load is 1029g / L filler, the sample loading is compl...

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 the technical field of antibody purification, in particular to a method for purifying antibody through cation exchange chromatography. The method comprises the following steps: performing cation exchange chromatography treatment on an antibody solution, adjusting the pH of the antibody solution to be 5.0 to 7.0 and the conductivity to be 3 to 20mS / cm, performing sample loading, adjusting the pH of an equilibration buffer to be 5.0 to 7.0 and the conductivity to be 3 to 20mS / cm, and performing purification through a flowthrough method or an overload method. The method has the advantages that the antibody is purified through the cation exchange chromatography, and the pH and conductivity of the antibody solution and the equilibration buffer are adjusted, so that target antibody monomer cannot be or is rarely combined with a cation chromatographic column or membrane, and flows out with flowthrough liquid, and impurity components can be combined with the chromatographic column or membrane, so that the purpose of antibody purification is achieved; meanwhile, the antibody yield can reach greater than 90 percent.

Description

technical field [0001] The invention relates to the technical field of antibody purification, in particular to a method for purifying antibodies by cation exchange chromatography. Background technique [0002] At present, there are hundreds of monoclonal antibody drugs on the market and in the research and development stage, and people's interest in biopharmaceuticals is also increasing. The characteristics of low toxicity and side effects, high specific therapeutic efficacy, long half-life and platform-based production technology make monoclonal antibody a leading position in the field of therapeutic biological products, and it is the most successful type of biological products. So far, most monoclonal antibodies are expressed by mammalian cells, which have high homology and similar molecular characteristics. These similar characteristics have been developed into a platform technology by many companies and research institutions to achieve efficient purification of a large ...

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): C07K16/00C07K1/18
CPCC07K1/18C07K16/00
Inventor 方峰梁泊宁汤炜周佳豪
Owner HJB HANGZHOU CO LTD
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