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Development of a shake tube‐based scale‐down model for perfusion cultures
- Source :
- Biotechnology and Bioengineering. 115:2703-2713
- Publication Year :
- 2018
- Publisher :
- Wiley, 2018.
-
Abstract
- The use of benchtop bioreactors (BRs) for the development of mammalian cell perfusion cultures is expensive and time consuming, given its complexity in equipment and operation. Scale-down models, going from liter to milliliter scale, are needed to support the rapid determination of suitable operating conditions in terms of viable cell density (VCD), perfusion rate, and medium composition. In this study, we compare the performance of steady-state perfusion cultures in orbitally shaken tube and BR systems for a given Chinese hamster ovary cell line. The developed scale-down model relied on a daily workflow designed to keep the VCD constant at specific target values. This includes: cell count, removal of excessive cells (bleeding), spin down of remaining cells, harvest of cell-free supernatant, and resuspension in fresh medium. Steady-state cultures at different VCD values, medium exchange rates and working volumes were evaluated. Shake-tube perfusion cultures allowed the prediction of cell-specific growth, glucose consumption, ammonia, and monoclonal antibody production rates for much larger BRs, but not lactate (LAC) production rates. Although charge variant profiles remained comparable, different glycosylation patterns were obtained. The differences in LAC production and glycosylation probably resulted from the discontinuous medium exchange, the poor carbon dioxide removal, and the deficient pH control. Therefore, if requested by the specific process to be developed, product quality has to be fine-tuned directly in the BR system. Altogether, the developed strategy provides a useful scale-down model for the design and optimization of perfusion cultures with strong savings in time and media consumption.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Cell Survival
Cell Culture Techniques
Cell Count
Bioengineering
CHO Cells
Shake
01 natural sciences
Applied Microbiology and Biotechnology
03 medical and health sciences
Bioreactors
Cricetulus
Perfusion Culture
010608 biotechnology
Bioreactor
Animals
Chromatography
Chemistry
Chinese hamster ovary cell
Hydrogen-Ion Concentration
Culture Media
030104 developmental biology
Perfusion rate
Cell culture
Perfusion
Scale down
Biotechnology
Subjects
Details
- ISSN :
- 10970290 and 00063592
- Volume :
- 115
- Database :
- OpenAIRE
- Journal :
- Biotechnology and Bioengineering
- Accession number :
- edsair.doi.dedup.....5911722f45552e11049412f4a56d4106