Chemical component-limited high-efficiency fed-batch culture medium, and preparation method and application thereof

A feed medium and chemical composition technology, applied in the field of feed medium, can solve the problems of cell density and antibody protein expression that need to be further improved, difficulty in preparing high-concentration products, and precipitation of nutrients, so as to reduce the dilution effect, Low production cost and the effect of avoiding precipitation

Active Publication Date: 2020-10-09
上海多宁生物科技股份有限公司
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  • Abstract
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Problems solved by technology

However, the feed medium contains alkali-soluble amino acids, which have low solubility at neutral pH, and it is difficult to prepare high-concentration products. In addition, nutrients will precipitate when stored for a long time or stored at low temperature, and the stability of the product is average, and CHO cell culture The final cell density and antibody protein expression still need to be further improved

Method used

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  • Chemical component-limited high-efficiency fed-batch culture medium, and preparation method and application thereof
  • Chemical component-limited high-efficiency fed-batch culture medium, and preparation method and application thereof
  • Chemical component-limited high-efficiency fed-batch culture medium, and preparation method and application thereof

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Embodiment 1

[0058] Select the optimal formula composition and content of the present invention to prepare, wherein the composition and formula of Duo Ning feeding medium A:

[0059]

[0060]

[0061] The method of preparing Donin feed medium A is as follows:

[0062] (1) Quantitatively weigh each component in the feed medium according to the prepared volume, and mix well;

[0063] (2) dissolving the feed medium mixture in step (1) in ultrapure water, controlling the water temperature at 20-30° C., and adding sodium hydroxide to adjust the pH to promote dissolution;

[0064] (3) Add 75g of glucose in every liter of culture medium and stir evenly;

[0065] (4) adjust the pH between 6.7-6.9, and use ultrapure water to make it volume;

[0066] (5) Filter with a 0.22 μm filter membrane and store at 2-8°C.

[0067] Composition and formula of Donin feed medium B:

[0068] amino acid Contentmg / L amino acid Contentmg / L L-tryptophan 28000 L-cysteine 24400 L-tyr...

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Abstract

The invention relates to the technical field of fed-batch culture media. The invention particularly discloses a chemical component-limited high-efficiency fed-batch culture medium. The culture mediumconsists of a feeding culture medium A and a feeding culture medium B, which are immiscible; wherein the feeding culture medium A contains amino acid with the total concentration of 12900 to 124000 mg/L; wherein the total concentration of the inorganic salt and the trace elements is 1210-12560 mg/mL, the total concentration of the vitamins is 780-10260 mg/L, and the total concentration of the other components is 1300-8020 mg/mL; and the feeding culture medium B contains amino acid with the total concentration of 18000 mg/L to 180000 mg/L. The fed-batch culture medium is high in nutrient concentration, especially high in amino acid and vitamin content, so that the dilution effect can be reduced, the cell density and antibody protein expression can be improved, and compared with other commercial fed-batch culture media, the fed-batch culture medium is good in culture effect, low in production cost and obvious in advantage.

Description

technical field [0001] The invention relates to the technical field of feed medium, in particular to a high-efficiency feed medium with defined chemical components, a preparation method of the feed medium, and an application of the feed medium in CHO cell culture. Background technique [0002] Chinese hamster ovary (CHO) cells are the most widely used host cells in the biomedical industry because they have post-translational modification capabilities similar to those of humans and can grow in high-density suspension, making them suitable for large-scale culture. Cells are very sensitive and fragile after isolation, so a suitable environment for their growth must be provided. The culture medium is a simulation of the in vivo environment, which is rich in nutrients and can meet the normal growth and metabolism of cells. [0003] Medium can be divided into basal medium and feed medium. Cells are first inoculated into the basal medium. As the cells grow and metabolize, the nut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
CPCC12N5/0682C12N2500/10C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/22C12N2500/24C12N2500/30C12N2500/32C12N2500/36C12N2500/38C12N2510/02
Inventor 王龙纪明宇王猛
Owner 上海多宁生物科技股份有限公司
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