Supercharge Your Innovation With Domain-Expert AI Agents!

A method for determining the loading amount of continuous flow chromatography and its application

A definite method, flow chromatography technology, applied in the direction of peptide preparation methods, chemical instruments and methods, instruments, etc., can solve the problems such as the inability to determine the sample volume, achieve faster data processing speed, greater convenience, and take into account the yield and the effect of filler loading

Active Publication Date: 2019-09-27
HJB HANGZHOU CO LTD
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The first purpose of the present invention is to provide a method for determining the loading amount of continuous flow chromatography, so as to solve the technical problem that the appropriate loading amount cannot be determined in continuous flow chromatography existing in the prior art

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 method for determining the loading amount of continuous flow chromatography and its application
  • A method for determining the loading amount of continuous flow chromatography and its application
  • A method for determining the loading amount of continuous flow chromatography and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This example provides a method for determining the loading amount of monoclonal antibodies expressed by CHO cells separated and purified by continuous flow chromatography. In the crude monoclonal antibody expressed by CHO cells to be purified, the expression amount is 3 g / L The pH of the sample was 7.0, and the conductivity was 15 mS / cm.

[0052] The method for determining the loading amount of the present embodiment includes the following steps:

[0053] (1) Take 1 g of monoclonal antibody protein expressed by CHO cells purified by single-column chromatography, adjust the pH to 7.0, conductance 15 mS / cm, and use equilibrium solution (50 mM NaAc-HAc, 150 mM NaCl, pH 7.4 for protein concentration) ) was diluted to 3g / L, and two columns were applied in series; wherein, the filler of the chromatography column was Mabselect Sure LX of GE, the column volume was 5mL (1.1cm*5.3cm), and the retention time was 2min;

[0054] (2) adopt the equilibration solution of 5CV (50mM NaA...

Embodiment 2

[0062] This example provides a method for determining the loading amount of monoclonal antibodies expressed by CHO cells separated and purified by continuous flow chromatography. In the crude monoclonal antibody expressed by CHO cells to be purified, the expression amount is 3.3 g / L , the loading pH is 7.2, and the conductivity is 15 mS / cm.

[0063] The method for determining the loading amount of the present embodiment includes the following steps:

[0064] (1) Take 1 g of monoclonal antibody protein expressed by CHO cells purified by single-column chromatography, adjust the pH to 7.2, conductance 15 mS / cm, and use equilibrium solution (50 mM NaAc-HAc, 150 mM NaCl, pH 7.4 for protein concentration) ) was diluted to 3.3g / L, and one column was used to drag two columns in series for sample loading; wherein, the chromatography column was HiTrap MabSelect SuRe 1mL prepacked column of GE Company, and the retention time was 2min;

[0065] (2) adopt the equilibration solution of 5CV (...

Embodiment 3

[0073] This example provides a method for determining the loading amount of monoclonal antibodies expressed by CHO cells separated and purified by continuous flow chromatography. In the crude monoclonal antibody expressed by CHO cells to be purified, the expression amount is 0.5 g / L , the loading pH was 7.15, and the conductance was 14 mS / cm.

[0074] The method for determining the loading amount of the present embodiment includes the following steps:

[0075] (1) Take 1 g of monoclonal antibody protein expressed by CHO cells purified by single-column chromatography, adjust the pH to 7.15, conductance 14 mS / cm, and use equilibrium solution (50 mM NaAc-HAc, 150 mM NaCl, pH 7.4 for protein concentration) ) was diluted to 0.5g / L, and two columns were dragged in series for loading; wherein, the packing of the chromatography column was Eshmuno A of Merck Company, the column volume was 5mL (1.1cm*5.3cm), and the retention time was 2min;

[0076] (2) adopt the equilibration solution...

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, and particularly relates to a determining method of sample loading amount chromatography through a continuous flow. The method comprises the steps that a target protein pure product is produced into sample liquid, and sample loading is conducted to obtain a penetrating curve; protein penetrating amount is calculated according to the formula to obtain DBC corresponding to the data point of C / C0=a in the penetrating curve to be taken as penetrating loading amount b; X corresponding to the data point where the protein penetrating amount Pn=100b+ / - 10 or Pn=200B+ / -10, and the sample loading amount is gamma(X-b) or gamma(X-2b), wherein a<=5% and 0.8<=gamma<=1. the method has the advantages that the sample loading amount under different conditions can be accurately given to take yield and filler loading amount into consideration at the same time, the method can be applied to the separation and purification of various kinds of proteins and the like, the determining method is simple, software of integral processing and the like is not needed, the method is convenient, the operation is convenient, and the learning of themethod is easy.

Description

technical field [0001] The present invention relates to the technical field of antibody purification, in particular to a method and application for determining the loading amount of continuous flow chromatography. Background technique [0002] With the advancement of biopharmaceutical technology, especially the development of upstream cell fermentation technology, the expression level of antibodies has gradually increased, from a few tenths of a gram per liter to as much as 30 grams per liter now, which brings great benefits to downstream purification. great pressure. The biggest barriers currently constraining downstream purification are the high cost of media for the capture step and the low production efficiency. The crude purification step of antibodies is currently mainly captured by Protein A affinity chromatography, and the widely used Protein A fillers generally have problems such as high price and low loading. [0003] The biopharmaceutical community has proposed ...

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): G16B25/00G16B40/00C07K1/16
Inventor 张赶富敏霞梁泊宁
Owner HJB HANGZHOU CO LTD
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com