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Method for syncretizing protein employing composite chromatography medium purification

A fusion protein and composite layer technology, which is applied in the field of biological protein purification, can solve the problems of large load impact and unsuitability for large-scale production, and achieve the effects of clear process control indicators, improved quality of final products, and tight process connection

Inactive Publication Date: 2008-12-10
陈志南 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, its loading capacity is greatly affected by the sample processing effect and operating conditions, so it is not suitable for large-scale production

Method used

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  • Method for syncretizing protein employing composite chromatography medium purification
  • Method for syncretizing protein employing composite chromatography medium purification
  • Method for syncretizing protein employing composite chromatography medium purification

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experiment example

[0082] 1. Cell solution clarification

[0083] Using 0.22 μm capsule filter (PALL company), silica gel tube, peristaltic pump (Baoding Lange Pump Co., Ltd.), filter the CHO cell culture supernatant at a constant flow rate to remove cells and cell debris.

[0084] 2. MabSelect SuRe affinity chromatography

[0085] Using the AKTA purifier-100 (GE Healthcare) chromatography system, 20ml of MabSelect SuRe (GE Healthcare) affinity chromatography medium was loaded into the XK16 / 20 (GE Healthcare) chromatography column. Fully equilibrate the affinity chromatography column with equilibration buffer (20mM PB+150mM NaCl pH 6.0-7.0), wait for the UV absorption at 280nm to return to the baseline, and start loading the sample when the conductance and pH remain stable, and then wash the remaining column with equilibration buffer. Binding protein, after the 280nm UV absorption returned to the baseline and the conductance and pH remained stable, it was directly eluted with elution buffer (10...

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Abstract

The invention discloses a method for adopting a compound chromatography medium to purify Fc fusion protein. The method comprises the following steps that: the ion-exchange hydrophobic compound chromatography medium is adopted to adjust the pH of a chromatography buffer system to be below the average pI of purification target protein and to adjust the chromatography buffer system to have high ionic strength simultaneously, utilizes proper chromatography conditions to ensure that the purification target protein exists in flow-through liquid; the flow-through liquid is collected, while polluting impurities (such as host cell protein, polymer, etc.) are adsorbed onto the compound chromatography medium, so as to improve the purity of the target protein. The method ensures that: the removal rate of final impurities reaches over 1 percent; the purity of the target protein reaches over 99 percent; the recovery rate of the target protein reaches over 90 percent. The method is compact in cohesion in realization process, can effectively remove the polluting impurities in one step, and is characterized in good stability, clear in process control index, suitable for mass production, etc.

Description

technical field [0001] The invention belongs to the field of biological protein purification, and relates to the purification of antibody proteins, in particular to a method for purifying Fc fusion proteins by using a composite chromatographic medium. The method is mainly used for the production and preparation of therapeutic Fc fusion protein medicine produced by large-scale culture of animal cells. Background technique [0002] Antibody therapy has become an important means of biological treatment of tumors, epidemics and infectious diseases today. Antibody drugs have become a large class of new therapeutic agents in the international pharmaceutical market due to their high specificity, effectiveness and safety. [1] . [0003] Large-scale cultivation of animal cells to produce antibody drugs has become the mainstream in the field of biopharmaceuticals. Many clinical antibody drugs, especially therapeutic drugs, require higher doses or long-term medication, and the marke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/18C07K1/20
Inventor 米力唐浩王科伟
Owner 陈志南
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