Serum-free high density suspension perfusion culture technology of hybridoma cells

A hybridoma cell and perfusion culture technology, which is applied in the field of hybridoma cell serum-free high-density suspension perfusion culture technology, can solve the problems of difficult to stabilize the culture state and increased substrate consumption, and achieve easy operation, small batch-to-batch difference, Reproducible and continuous effects

Active Publication Date: 2012-03-28
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

However, under high-density culture conditions, the substrate con

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  • Serum-free high density suspension perfusion culture technology of hybridoma cells
  • Serum-free high density suspension perfusion culture technology of hybridoma cells
  • Serum-free high density suspension perfusion culture technology of hybridoma cells

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Embodiment Construction

[0051] In view of the existing conditions and previous experience in large-scale cell culture, the applicant optimized the CSPR perfusion culture model and applied it to a hybridoma cell line (CYL-1 colon cancer antibody-expressing cell line). The design idea is to adjust the concentration of the main substrates (glucose and glutamine) in the perfusion medium on the basis of perfusion control. By-products accumulate rapidly. On the other hand, through the related control of perfusion rate and substrate flow rate, the loss of cells and products caused by a large amount of perfusion basal medium is avoided.

[0052] In the specific calculation process, the main control parameters considered are the perfusion rate and the substrate concentration of the perfusion medium, and the substrate concentration is controlled by the amount of concentrated medium added. According to the theory of reaction kinetics, the cell culture process can be divided into three stages, namely logarithmic...

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Abstract

The invention discloses a technology for producing a recombinant antibody protein through the high density perfusion culture of hybridoma cells. The technology which aims at the metabolic characteristic of the hybridoma cells screens a specific culture component for the perfusion culture, and allows metabolic substrates to be fully supplied in the culture process and the concentration of metabolic byproducts to be simultaneously and effectively reduced, so objects of high efficiency and high yield is reached. In the process control aspect, a specific perfusion rate CSPR perfusion culture calculating model is established to carry out real time calculation on culture parameters, and concentrated medium subsection fed batch is adopted to control the perfusion rate, so important substrates ofglucose and glutamine are supplied and the influence of the byproducts to culture is minimized. The technology which is suitable for the preparation of the production antibody protein through the large scale culture of the hybridoma cells allows a large amount of protein antibodies to be provided in a short time, has the advantages of small scale, low investment, realization of the high efficiency output in a short time, low batch difference, high repeatability, and realization of the production of antibody medicines.

Description

technical field [0001] The invention belongs to the cell engineering technology in the field of cell engineering, and mainly relates to the large-scale cultivation of hybridoma cells and the production of recombinant protein antibodies, in particular to a serum-free high-density suspension perfusion culture process of hybridoma cells. Background technique [0002] Animal cell culture process is the production process of animal cells expressing biotechnology products. The research of this technology is to use the biological metabolic process of animal cells to control it and solve the commonality and characteristics of the process. The main subject bases of this process research are cell biology and biochemical engineering. Related disciplines include genetics, immunology, molecular biology, biochemistry, microbiology, chemical engineering, applied physics and electronic computers. It is an emerging biotechnology composed of multidisciplinary basic synthesis. Therefore, pe...

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Application Information

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IPC IPC(8): C12N5/20C12P21/08C12R1/91
Inventor 王彬张征张阳屈颖南刚孙秀璇徐力清姚西英王丽娟
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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