Method for culturing cells by using circular-track shaking type bioreactor

A bioreactor and cell culture technology, which is applied in the field of culturing cells using an orbital shaking bioreactor, can solve problems such as differences in working principles and unsuitable culture conditions, and achieve improved expression, high protein expression and specific production. speed, the effect of prolonging the incubation time

Pending Publication Date: 2021-12-31
CANTONBIO CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is a big difference between the working principle of the orbital shaking bioreactor and the stirred b

Method used

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  • Method for culturing cells by using circular-track shaking type bioreactor
  • Method for culturing cells by using circular-track shaking type bioreactor
  • Method for culturing cells by using circular-track shaking type bioreactor

Examples

Experimental program
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Example Embodiment

[0054] Example 1

[0055] Using cyclic rail shaken bioreactor cell culture method, in a certain cell line cultured cells are seeded in a culture medium. 1, Comparative Example 1, as shown in Table 1 Example 2 Comparative cell culture conditions embodiments.

[0056] Culture Conditions Table 1 Comparative Example of

[0057]

[0058]

[0059] Note: N / a means it does not apply.

[0060] Real-time monitoring of the content of glucose in the medium during culture and glutamine, as its content is less than the minimum value of the defined range, it is necessary to add glucose and glutamine in the medium.

[0061] pH Control Strategy: Real-time monitoring of medium pH, if the pH is lower than the above pH range, the addition of 1M NaHCO 3 Elevated pH, if the pH is below the above-described pH range, is passed into CO 2 Lower pH.

[0062] DO Control Strategy: Real-time monitoring of dissolved oxygen in the medium, the dissolved oxygen into the air within a defined range.

[0063] Ce...

Example Embodiment

[0071] Example 2

[0072] In addition to external pH of the medium in Example 2 and Example 1 are different, the other identical culture conditions. Example embodiment 2 medium pH was 7.0.

[0073] Cell culture process, cell density is detected using a cell counter (Beckman), survival rate, using the biochemical analyzer (U.S. NOVA) detecting nutrients. After completion of cell culture, the supernatant was collected after centrifugation. The supernatant by HPLC, collected anti-human CD20 monoclonal antibody (anti-CD20 Mab), to detect its expression yield.

[0074] The results showed that close to the fixed cell growth pH setpoint state in Example 1, the pCO 2 The proportion has increased, not decreased expression.

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Abstract

The invention discloses a method for culturing cells by using a circular-track shaking type bioreactor. The method is characterized in that a pH value and a rotating speed of cell culture conditions and other culture conditions are adjusted, and then, the circular-track shaking type bioreactor is used for culturing the cells. Results show that the cell culture method disclosed by the invention has no obvious difference in cell metabolism in a cell culture process, but a pH change range is smaller than that of a stirring type bioreactor, and damage to the cells is reduced; by the culture method provided by the invention, relatively high cell viability can be kept for a long time, and the culture time is prolonged; and an expression level curve and a specific production rate curve are relatively consistent, and the protein expression level can be relatively obviously increased by prolonging the culture time in the circular-track shaking type bioreactor.

Description

technical field [0001] The invention relates to the field of biotechnology, and in particular to a method for cultivating cells using a ring-track shaking bioreactor. Background technique [0002] Monoclonal antibodies play an important role in biopharmaceutical products. At present, monoclonal antibodies are mainly expressed and produced through in vitro culture of animal cells. The stirred bioreactor is a commonly used reactor in the field of cell culture. The technology is relatively mature, but there are still many problems, which bring obstacles to the process of cell scale-up culture. First of all, during the scale-up culture process, the shear force caused by the stirring paddle will cause damage to the cells, and even death. Secondly, the stirred bioreactor generally adopts the deep ventilation method, which is easy to generate foam, and the rupture of the bubble will cause a greater shear force. In addition, in the process of expanding the culture of stirred biore...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/13
CPCC12N5/0682C07K16/2887C07K16/32C12N2500/00C12N2500/60C12N2500/32C12N2500/34C12N2500/02C07K16/00C12N5/10C12M1/36C12N5/06C12M3/00
Inventor 景涛陆金康吴小波胡飞
Owner CANTONBIO CO LTD
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