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Animal cell suspension culture process utilizing riptide bioreactor, and controlling method thereof

A bioreactor and animal cell technology, applied in the field of cell engineering, can solve the problems of mass transfer, low oxygen transfer efficiency, poor mixing effect, etc.

Active Publication Date: 2013-03-27
哈药集团股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biggest disadvantage of this reactor is that the mixing effect is poor, and the efficiency of mass transfer and oxygen transfer is low.

Method used

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  • Animal cell suspension culture process utilizing riptide bioreactor, and controlling method thereof
  • Animal cell suspension culture process utilizing riptide bioreactor, and controlling method thereof
  • Animal cell suspension culture process utilizing riptide bioreactor, and controlling method thereof

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Embodiment Construction

[0025] The content of the present invention will be described in detail below through examples, but these examples do not constitute a limitation to the present invention. The following takes the large-scale culture of CHO cells (CHO cells expressing recombinant anti-EGFR human-mouse chimeric monoclonal antibody) as an example to illustrate the production process.

[0026] 1. Preparation of culture medium

[0027] 1.1.1 Preparation of basal culture medium:

[0028] Take normal temperature water for injection, first add about 70% of the total volume of the preparation. Accurately weigh the required culture medium, add CDM4CHO dry powder (6.25 g / L) from HyClone Company and stir for 30 minutes. Add soybean hydrolyzate 1g / L, yeast hydrolyzate 3g / L and continue stirring for 30 minutes. Add the following substances: KCl (0.3118g / L), NaH 2 PO 4 (0.125g / L), Na 2 HPO 4 (0.142g / L)NaHCO 3 (1.5g / L), HEPES (4.76g / L), F68 (1.5g / L), NaCl (2.5g / L), Glucose (2.5g / L), Glutamine (0.585g / ...

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Abstract

The invention discloses a process of serum-free culture of animal cells for producing recombinant protein drugs by using a riptide bioreactor, and a controlling method thereof. According to the process, through basic culture medium feeding, cell density is improved, and cell culture volume is enlarged. Through reducing a culture temperature and feeding a concentrated expression culture medium, cell density and survival rate are maintained, such that recombinant protein can be highly efficiently and continuously obtained. According to the process controlling method, on-line monitoring is carried out upon pH values, dissolved oxygen, and temperature changes aiming at growth and metabolism changes of different cell lines; and timed detections are carried out upon cell density, survival rate, glucose in the culture system, glutamine, ammonia, lactic acid, and osmotic pressure. The method is especially suitable for large-scale animal cell suspension culture. The process has the advantages of simple operation and controlling, high mass transfer and oxygen transfer efficiency, stable culture period, small difference between batches, and low production cost. With the process, large volume and high density are easy to realize.

Description

technical field [0001] The invention belongs to cell engineering technology, and relates to large-scale cultivation of animal cells and a process for producing recombinant protein drugs, in particular to a process control method for animal cell serum-free high-density, high-expression suspension culture. Background technique [0002] Large-scale cultivation of animal cells has become one of the most important key technologies in the field of biopharmaceuticals, and has developed rapidly with the wide application of recombinant therapeutic protein drugs. At present, the main method of industrial production of recombinant protein therapeutic drugs expressed by animal cells is to use bioreactors to culture hybridoma cells, engineered cells, insect cells and microorganisms in large-scale and high-density in vitro, and then prepare recombinant protein products through related purification methods. [0003] Bioreactors for large-scale cell culture include stirred bioreactors, airl...

Claims

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Application Information

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IPC IPC(8): C12N5/02C12N5/07C12P21/00C12P21/08C12M3/02C12M1/38C12M1/36
Inventor 张秀芹刘铁成张雪亭王延涛杨威李会成
Owner 哈药集团股份有限公司