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Cell culture method for reducing acidic peak content of antibody and improving glycoform of antibody

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: 2016-07-20
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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  • Cell culture method for reducing acidic peak content of antibody and improving glycoform of antibody
  • Cell culture method for reducing acidic peak content of antibody and improving glycoform of antibody
  • Cell culture method for reducing acidic peak content of antibody and improving glycoform of antibody

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Experimental program
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Effect test

Embodiment 1

[0033] Culture of a cell line A expressing anti-tumor necrosis factor-α (TNFα) antibody.

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

[0035] The cell culture process involves detection of cell viability and density, concentra...

Embodiment 2

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

[0052] In the cell culture experiment, on the 0th day, the basal medium CDFortiCHOAGT with the final concentrations of 0mM (control group), 2mM, 4mM, and 6mM glutamine were all added to 500ml shake flasks, and the cells were divided into 1×10 6 Cells / mL was inoculated and cultured at 37°C. From the third day, the cell viability and density and various biochemical parameters in the culture were measured every day, and fed-batch culture was carried out. The fed-batch medium was EfficientFeed TM B+, according to the results detected by the biochemical analyzer, the glucose concentration was controlled at 10g / L, the monosodium glutamic acid concentration was 8mM, the temperature was controlled at 34°C from the sixth day, and the cells were harvested on the 13th day of cell culture. The antibody quality detection method is the sam...

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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 antibodies and improving antibody glycoforms. Background technique [0002] CHO cells (Chinese Hamster Ovary 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 post-translational modifications. The protein structure, immunogenicity, glycosylation type and method, etc. are almost the same as those of the natural protein, so that the drug and efficacy of the recombinant protein can be guaranteed. In addition, CHO cells rarely secrete their own endogenous proteins, which is also conducive to the later isolation and purification of exogenous proteins. [0003] Monoclonal antibodies have a complex molecular structure and a large molecular weig...

Claims

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Application Information

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Patent Type & Authority Applications(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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