CHO cell culture method in bioreactor

A bioreactor, cell culture technology, applied in the field of CHO cell culture in bioreactors, can solve the problems of no obvious improvement in antibody yield and quality, poor versatility, etc., to achieve wide applicability, reduce costs, and shorten the cycle. Effect

Inactive Publication Date: 2019-09-17
君科华仞(北京)医药科技有限公司
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Problems solved by technology

[0003] In some existing literatures, it is reported that certain specific rotation speed, temperature, pH and dissolved oxygen control will affect the growth of cells and the quality of antibodies, but these studies often only consider the influence of a single factor. Cultivated in shake flasks, the versatility is poor, and there is no significant improvement in the yield and quality of antibodies

Method used

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  • CHO cell culture method in bioreactor
  • CHO cell culture method in bioreactor
  • CHO cell culture method in bioreactor

Examples

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Embodiment 1

[0039] The CHO cell culture process of the present embodiment is as follows:

[0040] (1) Insert the basal medium into the reactor, inoculate two GS-CHO antibody expressing cell lines A and B respectively, the inoculation density is 0.6106cells / ml, set the dissolved oxygen to 40%, the temperature is controlled at 37C, the rotation speed is 140rpm, and CO2 and NaHCO3 solution to adjust the pH to 7.0-7.3;

[0041] (2) After culturing for a certain period of time (3rd day), control the pH range to 6.8 to 7.0, and adjust the rotation speed to 220rpm on the 6th day;

[0042] (3) From the 4th day, feed once every 2 days, the volume of the feed medium added each time is 5% of the initial culture volume, and stop feeding when the cell viability drops below 80%;

[0043] (4) The basic medium is Dynamis, and the fed-batch medium is EfficientFeedTM C+;

[0044] (5) Shake flask culture is a control group, and the shake flask culture conditions are: temperature control 37°C, CO Concentra...

Embodiment 2

[0049] This example adopts the same method as Example 1 for culturing, wherein the basal medium is ActiPro, the fed-batch medium is Cell Boost 7a / 7b, and the inoculated cell line is GS-CHO antibody-expressing cell line C. This example also uses Shake flask culture was used as control group.

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Abstract

The invention discloses a CHO cell culture method in a bioreactor. The key point of the technical scheme comprises the following steps: (1) initially controlling a bioreactor; (2) controlling the pH value in the early stage of culture; (3) adjusting the pH value in the medium period of culture and simultaneously controlling the stirring speed of the bioreactor; (4) inoculating the stirred material with basic culture medium in the process of the step (2); and (5) adding the fed-batch culture medium in the process of the step (3). The method has wide applicability, and can obviously reduce the cost and shorten the period.

Description

technical field [0001] The present invention relates to a kind of cell culture, more specifically, it relates to a kind of CHO cell culture method in the bioreactor. Background technique [0002] Antibody drugs have become leaders in the field of biopharmaceuticals due to their advantages such as clear targeting and small side effects. Most of the new antibody drugs currently on the market are produced on the basis of large-scale CHO cell culture. In order to ensure that the expression quantity and quality of antibodies meet the requirements, the cultivation process in the bioreactor also needs to be screened and optimized. Due to the enlarged volume, the research and development costs at this stage will increase significantly. In the early stage of research and development of antibody drugs, a large amount of screening and process development work is mostly carried out in well plates, shake flasks or shake tubes. Due to different culture modes, how to quickly transition th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12Q3/00
CPCC12N5/0682C12Q3/00
Inventor 郝学才李文宾
Owner 君科华仞(北京)医药科技有限公司
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