Serum-free protein-free feed culture medium as well as preparation method and application thereof

Active Publication Date: 2017-12-12
上海多宁生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, there is a lot of blindness in the selection of suitable commercial feed medium, and when adjusting the medium composition to improve protein quality ind

Method used

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  • Serum-free protein-free feed culture medium as well as preparation method and application thereof
  • Serum-free protein-free feed culture medium as well as preparation method and application thereof
  • Serum-free protein-free feed culture medium as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0060] The preparation method (1L) of high-efficiency feeding medium of the present invention

[0061] Prepare according to the composition ratio of the high-efficiency feed medium of the present invention (see Table 1)

[0062] Add 800ml of ultra-pure water at a temperature of about 35°C, weigh each component according to the composition ratio of the high-efficiency feeding medium of the present invention, add a certain amount of sodium hydroxide to promote dissolution, and then add concentrated hydrochloric acid to adjust the pH to 6.9-7.2 In between, filter with a 0.2 μm filter membrane, and store in 1L glass bottles aseptically protected from light at 4°C.

[0063] Table 1: Composition (formula one) and ratio of the high-efficiency feed medium of the present invention

[0064]

[0065]

[0066]

Embodiment 2

[0068] According to the configuration of Example 1, other feed medium ratios are provided.

[0069]

[0070]

Embodiment 3

[0072] Fed-fed medium (supplemented feed medium) to CHO cell culture medium

[0073] The Hyclone feed medium used in the embodiments of the present invention is purchased from GE Healthcare Hyclone, and is Cell Boost 2 (catalogue number SH30596), Cell Boost 4 (catalogue number SH30857), Cell Boost 5 (catalogue number SH30865), Cell Boost 6 ( Cat. No. SH30866); GIBCO feed medium was purchased from Thermo Fisher Invitrogen as Feed A+ (Cat. No. A25023), Feed B+ (Cat. No. A25030), Feed C+ (Cat. No. A25031), Feed C (Cat. No. A13275), TiterEnhancer ( Cat. No. A15010); Irvine BalanCD CHOfeed feed medium was purchased from Irvine Scientific as Feed 1 (Cat. No. 94119), Feed 2 (Cat. No. 94121), Feed 3+ (Cat. No. 94118); EX-CELL Advanced TM CHO Feed 1 was purchased from Sigma with catalog number 24367C; CellventoFeed-210 was purchased from Merck with catalog number 102488; Kerry Sheff-CHO hydrolyzate feed was purchased from Cimege as ACF (catalog number 5×00424), PF ACF (Cat. No. 5x004...

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Abstract

The invention relates to a serum-free protein-free high-efficiency feed culture medium as well as a preparation method and application thereof. The feed culture medium comprises amino acids, inorganic salts, trace elements, vitamins, carbohydrates and other organic matters. Moreover, the culture medium does not contain any glutamine. Since the feed culture medium contains serum-free protein-free animal-source-free components, the viral pollution risk can be greatly reduced, and downstream purification is facilitated. Meanwhile, since the culture medium is full in types of formula components and balanced in proportion, the culture medium can be applicable to high-density culture and high-protein expression of multiple CHO cell strains.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, the invention relates to a serum-free, protein-free high-efficiency feed medium and a preparation method and application of the medium. Background technique [0002] At present, CHO cells, as the main host cells, are widely used in the production of therapeutic protein drugs. The development of a high-yield, efficient, robust, low-cost, and high-quality production process is critical for the industrial production of therapeutic protein products. The key upstream technology platforms include: high-affinity fully humanized antibody screening platform, high-expression engineered cell line construction and screening platform, personalized high-efficiency basal medium and feed medium development platform, supporting high-density cell growth and High-expression culture process development platform, etc., among which low-cost and high-efficiency medium development is crucial for the indu...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/10C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/22C12N2500/30C12N2500/32C12N2500/34C12N2500/35C12N2500/36C12N2500/38C12N2500/46C12N2500/95C12N2501/999
Inventor 王龙王猛秦孙星陈智旺
Owner 上海多宁生物科技股份有限公司
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