Domesticated CHO-S cell system and culture method and application thereof

A CHO-S, cell line technology, applied in the field of bioengineering, can solve the problems of increasing product production costs, lack of core technology, scale risks, etc., and achieve the effects of short doubling time, increased expression, and good cell morphology

Active Publication Date: 2017-01-04
QILU PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although after years of development, my country's technical level in protein recombinant expression has been significantly improved, but it still lacks independent core technologies. Most of the cell lines and culture media used by enterprises or scientific research institutions are commercial products. These commercial products Not only will it significantly increase the production cost of future products, but it will also bring great risks to scale

Method used

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  • Domesticated CHO-S cell system and culture method and application thereof
  • Domesticated CHO-S cell system and culture method and application thereof
  • Domesticated CHO-S cell system and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1 Preparation of the domesticated CHO-S cell line of the present invention:

[0060] (1) Add Anti-Clumping Agent (0.1-1% by volume) and L-Glutamine (0.8-8mM) to CD FortiCHO medium, and preheat at 37°C for about 30 minutes; Take it out of liquid nitrogen, put it into a 37°C water bath to thaw quickly, and add it to the preheated CD FortiCHO medium in an ultra-clean bench, count with a CountStar cell counter, and control the inoculation density at 0.5-1.0×10 6 Cells / mL, at 37°C, 85% humidity, 5% CO 2 , 110-150rpm shaking flask culture;

[0061] (2) After the CHO-S cell line grows stably in the CD Forti CHO medium, select the cell line in the logarithmic growth phase that recovers better in the CD FortiCHO medium, and use 0.5×10 6 The cells / mL density were inoculated in XH001 medium and cultured in 125mL culture flasks;

[0062] (3) When the cell density is >2×10 6 Cells / mL density, again at 0.5×10 6 The cells / mL density were seeded in XH001 medium and cult...

Embodiment 2

[0068] Example 2: Comparative test on the growth of CHO-S cells of the present invention and commercialized CHO-S cells

[0069] After the acclimated cells obtained in step (2) of Example 1 were grown stably in the XH001 medium described in Example 1, the 6 The cells / mL cell density were respectively inoculated in the XH001 medium described in Example 1 for batch culture research. At the same time, the CD Fort iCHO medium was used as a control, and the CountStar cell counter was used to count the cell number and viability every 24 hours. cell growth curve image 3 shown.

Embodiment 3

[0070] Example 3: Evaluation of the stability of the CHO-S cell line of the present invention in XH001 medium

[0071] In the XH001 medium described in Example 1, the CHO-S cell strain was counted by the CountStar cell counter for cell number and survival rate every 24 hours, and every 3 days according to 0.5×10 6 cells / mL density for passage. After about 23 generations of growth stability evaluation, the growth of the cells was stable, and the doubling time was about 15 hours, indicating that the CHO-S cells adapted to the nutritional environment of the XH001 medium and grew well.

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Abstract

The invention relates to a domesticated CHO-S cell system and a culture method and application thereof. The domesticated CHO-S cell system is obtained by passing CHO-S cells for at least 5 generations through a specially formed XH001 culture medium by a gradual domestication method. The invention also discloses a domestication method and application of the domesticated CHO-S cell system. The domesticated CHO-S cell system has the characteristics of high growth speed, short multiplication time, good cell form, uniform shape and the like. When the domesticated CHO-S cell system is used for recombinant antibody expression, the expression quantity can be obviously improved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a domesticated CHO-S cell line and its culture method and application. Background technique [0002] With the continuous development of genetic engineering technology, recombinant biotechnology drugs have been developed rapidly, and a variety of eukaryotic and prokaryotic protein expression systems are used to express proteins with therapeutic value. Among these expression systems, the mammalian cell expression system has become the first choice for the recombinant expression of some medicinal proteins due to the high consistency of the expressed protein with the natural molecule in terms of molecular structure, post-translational modification and biological function. system. [0003] Since CHO (Chinese Hamster Ovary, Chinese Hamster Ovary) cells were successfully isolated, after decades of extensive basic research work, it has developed into the most importan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N1/36C12P21/00C12R1/91
Inventor 李剑凤杨建国朱蕾王桂江孙丽霞任志强范建民
Owner QILU PHARMA CO LTD
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