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New application of ATA in basal culture medium for culturing CHO cell line cells

A basic culture medium and cell line technology, applied in tissue culture, animal cells, reproductive tract cells, etc., can solve the problems of high price, high price of culture medium, restricting the development of biopharmaceutical industry, etc., and achieve the effect of reducing costs

Pending Publication Date: 2022-03-11
无锡多宁生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, IGF is an essential substance for some CHO cells, but the price is high, and the medium market in my country is currently monopolized by foreign companies such as Gibco and Hyclone, and the high price of the medium seriously restricts the development of the biopharmaceutical industry.

Method used

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  • New application of ATA in basal culture medium for culturing CHO cell line cells
  • New application of ATA in basal culture medium for culturing CHO cell line cells
  • New application of ATA in basal culture medium for culturing CHO cell line cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] According to the optimal concentration range of the guidance of the present invention, 5 concentration range points were selected respectively for the two kinds of substances to carry out the cell experiment. ), and the feeding medium Feed 2+feed B2 independently developed by Duoning was selected as the feeding material.

[0041] Test method: Fed-batch culture (adding feed medium)

[0042] Experimental procedure: Feed the cells in the middle of the logarithmic growth phase, and harvest them all on Day 13.

[0043] Culture environment: 37°C, 5% carbon dioxide

[0044] Shake flask culture process: Add the sterile-filtered basic medium to two 2L (No. 1 and No. 2) and one 125mL No. 3 shake flasks in the ultra-clean bench, and the inoculation density is 5×10 5 Cells / ml, the culture volume is 200ml, cultured to the 3rd day and counted: the viable cell density is 30.05×10 5 cells / ml, the cell viability was 98.36%. Evenly distribute 30mL culture medium of No. 1 1L shake fla...

Embodiment 2

[0051] The optimal concentrations of ATA and IGF were screened out through screening experiments, which were 100 mg / L and 0.2 mg / L, respectively (specific screening methods are not specifically described). Five different CHO cell lines (including CHOK-1) were selected and named as CHO-1, CHO-2, CHO-3, CHO-4, and CHO-5. The culture method is fed-batch culture (fed-batch), and the commercial basic medium (Media C) independently developed by Duo Ning is selected, and the feed medium Feed 2+feed B2 independently developed by Duo Ning is used as the feed.

[0052] Test method: Fed-batch culture (adding feed medium)

[0053] Experimental procedure: Feed the cells in the middle of the logarithmic growth phase, and harvest them all on Day 13.

[0054] Culture environment: 37°C, 5% carbon dioxide

[0055] Shake flask culture process: Add the sterile-filtered basic medium to 15 125mL shake flasks in the ultra-clean bench, and the inoculation density is 5×10 5 Cells / ml, the culture vo...

Embodiment 3

[0060] In order to determine the possibility of mutual substitution between ATA and IGF, this embodiment was carried out. CHO K-1 cells were selected, ATA and IGF were added together (concentrations were 50mg / L, 0.5mg / L and 150mg / L, 0.1mg / L), and 100mg / LATA and 0.2mg / LIGF were added separately. The culture method is fed-batch culture (fed-batch), and the commercial basic medium (Media C) independently developed by Duo Ning is selected, and the feed medium Feed 2+feed B2 independently developed by Duo Ning is used as the feed.

[0061] Test method: Fed-batch culture (adding feed medium)

[0062] Experimental procedure: Feed the cells in the middle of the logarithmic growth phase, and harvest them all on Day 13.

[0063] Culture environment: 37°C, 5% carbon dioxide

[0064] Shake flask culture process: Add the sterile-filtered basic medium to five 125mL shake flasks in the ultra-clean bench, and the inoculation density is 5×10 5 Cells / ml, the culture volume is 30ml, cultured ...

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Abstract

The invention discloses a novel application of ATA in a basic culture medium for culturing CHO cell line cells. A substance aurin tricarboxylic acid can be added into the CHO cell basic culture medium to promote cell growth and titer; aTA has a general promotion effect on different CHO cell lines, and it is found that the ATA has some similar modes with IGF in the aspect of a cell promotion mechanism, so that the ATA can be used as partial substitution or complete substitution of the IGF, and the cost of production raw materials is reduced on the premise that cell growth and expression are not affected; in addition, the ATA serving as a chemical raw material meets the requirement of serving as a chemically restricted culture medium, so that the ATA can be well applied to the production of recombinant proteins and vaccines, the production cost is reduced, and a good guarantee is provided for adapting to the industrial culture process in the future.

Description

technical field [0001] The invention relates to the technical field of biological cells, in particular to a new application of ATA in a basal medium for cultivating CHO cell line cells. 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 industrial production of therapeutic proteins. ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2501/999
Inventor 王龙杨一鸣王猛
Owner 无锡多宁生物科技有限公司
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