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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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  • Summary
  • 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 bioreactor, and the culture conditions of the stirred bioreactor are not suitable for the orbital shaking bioreactor.

Method used

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

Examples

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Effect test

Embodiment 1

[0055] A method for cultivating cells using a ring-orbit shaking bioreactor, inoculating cell strains in a medium at a certain density for cell cultivation. The conditions for culturing cells in Example 1, Comparative Example 1, and Comparative Example 2 are shown in Table 1.

[0056] The culture condition of table 1 comparative example and embodiment

[0057]

[0058]

[0059] Note: N / A means not applicable.

[0060] During the cultivation period, the contents of glucose and glutamine in the medium are monitored in real time, and if the contents are lower than the minimum value of the defined range, glucose and glutamine need to be added to the medium.

[0061] pH control strategy: monitor the pH of the medium in real time, if the pH is lower than the above pH range, add 1M NaHCO 3 Raise the pH, if the pH is below the above pH range, pass CO 2 Lower the pH.

[0062] DO control strategy: monitor the dissolved oxygen in the medium in real time, and control the dissolved...

Embodiment 2

[0072] Example 2 and Example 1 have the same culture conditions except that there is a difference in the pH value of the medium. The pH of the culture medium in Example 2 was 7.0.

[0073] During the process of culturing cells, a cell counter (Beckman Company) was used to detect cell density and viability, and a biochemical analyzer (NOVA, USA) was used to detect nutrients. After the cell culture was completed, the supernatant was collected after centrifugation. The supernatant was collected by high-performance liquid chromatography to collect anti-human CD20 monoclonal antibody (anti-CD20 Mab), and its expression yield was detected.

[0074] The results show that the fixed pH setting value cell growth state is close to that in Example 1, pCO 2 The proportion increased, but the expression level did not decrease significantly.

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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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