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A cell culture method for reducing antibody acidic peak content and improving antibody glycoform

A cell culture and antibody technology, applied in the field of cell culture, to achieve the effect of ensuring drug efficacy and optimizing glycosylation level

Inactive Publication Date: 2020-03-24
SUNSHINE LAKE PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these two peaks have similar chemical properties, and it is difficult to control the charge heterogeneity through purification and separation in the later stage, and it is also challenging to control the charge heterogeneity of antibodies by controlling the cell culture process, which has become a challenge in this field. Problems that have always been difficult to solve in the field

Method used

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  • A cell culture method for reducing antibody acidic peak content and improving antibody glycoform
  • A cell culture method for reducing antibody acidic peak content and improving antibody glycoform
  • A cell culture method for reducing antibody acidic peak content and improving antibody glycoform

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Cultivation of a cell line A expressing antibodies against tumor necrosis factor-α (TNFα).

[0034] In the cell culture experiment, on day 0, the basal medium CDM4PERMAb with final concentrations of 0mM (control group), 2mM, 4mM, and 6mM glutamine were added to 500ml shake flasks. The cells were divided into 1×10 6 Cells / mL were inoculated and cultured at 37°C. From day 3, cell viability and density and various biochemical parameters in the culture were measured every day, and fed culture was carried out. The fed culture medium was CHO CD Efficient Feed TM A. Control the glucose concentration at 5g / L and the monosodium glutamate concentration at 4mM according to the results of the biochemical analyzer, and control the temperature at 33°C on the 6th day, and harvest on the 15th day of cell culture.

[0035] The cell culture process involves the detection of cell viability and density, anti-TNFα antibody concentration, anti-TNFα antibody charge heterogeneity, anti-TNFα antibody...

Embodiment 2

[0051] Cell line B was cultured for expression of recombinant human tumor necrosis factor receptor-Fc recombinant protein (rhTNFR-Fc) antibody.

[0052] In the cell culture experiment, on day 0, the basal medium CD Forti CHO AGT with final concentrations of 0 mM (control group), 2 mM, 4 mM, and 6 mM glutamine were added to 500 ml shake flasks. 10 6 Cells / mL were inoculated and cultured at 37°C. From the third day on, the cell viability and density and various biochemical parameters in the culture were measured every day, and fed culture was carried out. The fed culture medium was Efficient Feed TM B+, according to the results of the biochemical analyzer, the glucose concentration was controlled at 10g / L, the monosodium glutamate concentration was at 8mM, the temperature was controlled at 34°C on the sixth day, and the cell culture was harvested on the 13th day. The antibody quality detection method is the same as in Example 1. The cell viability, viable cell density, antibody exp...

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Abstract

The invention discloses a cell culture method for reducing acidic peak content of an antibody and improving glycoform of the antibody. In the method, GS-CHO cells are used as an expression system, glutamine is added into a basic medium for antibody-expressed cell strain culture. After glutamine is added to the basic medium, it is possible to effectively reduce acidic peak content of the antibody and improve glycosylation level, thus modifying the activity and efficacy of antibody drugs and improving the quality of the antibody so that the quality is close to that of a standard product; the method is adapted to the development and study and later large-scale production of new antibody drugs and has a promising application prospect and good economic benefit.

Description

Technical field [0001] The invention relates to a cell culture method, in particular to a cell culture method for reducing the acidic peak content of an antibody and improving the glycoform of the antibody. Background technique [0002] CHO cells (Chinese Hamster Overy) are one of the most widely used host cells for the production of protein drugs. Compared with other cell expression systems, CHO cell expression systems can stably integrate with foreign genes and undergo complex translation. Modified and expressed proteins that have almost the same structure, immunogenicity, glycosylation type and method as the natural protein, so as to ensure the drug properties and efficacy of the recombinant protein. In addition, CHO cells rarely secrete their own endogenous proteins, which is also conducive to the later separation and purification of foreign proteins. [0003] Monoclonal antibodies have complex molecular structures and large molecular weights, and can produce a large number of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12P21/08
CPCC07K16/241C07K16/2878C07K2317/14C12N5/0682C12N2500/32
Inventor 高重亮杨彬鄢成伟黎美香阮永贤孙文正
Owner SUNSHINE LAKE PHARM CO LTD
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