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. 2020 Jul;117(7):1915-1928.
doi: 10.1002/bit.27335. Epub 2020 Apr 8.

Hydrocyclones as cell retention device for CHO perfusion processes in single-use bioreactors

Ioná W Bettinardi  1 , Andreas Castan  2 , Ricardo A Medronho  3 , Leda R Castilho  1
Affiliations

Hydrocyclones as cell retention device for CHO perfusion processes in single-use bioreactors

Ioná W Bettinardi et al. Biotechnol Bioeng. 2020 Jul.

Abstract

In this study, a hydrocyclone (HC) especially designed for mammalian cell separation was applied for the separation of Chinese hamster ovary cells. The effect of key features on the separation efficiency, such as type of pumphead in the peristaltic feed pump, use of an auxiliary pump to control the perfusate flow rate, and tubing size in the recirculation loop were evaluated in batch separation tests. Based on these preliminary batch tests, the HC was then integrated to 50-L disposable bioreactor bags. Three perfusion runs were performed, including one where perfusion was started from a low-viability late fed-batch culture, and viability was restored. The successive runs allowed optimization of the HC-bag configuration, and cultivations with 20-25 days duration at cell concentrations up to 50 × 106 cells/ml were performed. Separation efficiencies up to 96% were achieved at pressure drops up to 2.5 bar, with no issues of product retention. To our knowledge, this is the first report in literature of high cell densities obtained with a HC integrated to a disposable perfusion bioreactor.

Keywords: CHO cells; hydrocyclone; perfusion; separation efficiency; single-use bioreactor.

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References

REFERENCES

    1. Batt, B., Davis, R., & Kompala, D. (1990). Inclined sedimentation for selective retention of viable hybridomas in a continuous suspension bioreactor. Biotechnology Progress, 6(6), 458-464. https://doi.org/10.1021/bp00006a600
    1. Bhardwaj, P., Bagdi, P., & Sen, A. K. (2011). Microfluidic device based on a micro-hydrocyclone for particle-liquid separation. Lab on a Chip, 11(23), 4012. https://doi.org/10.1039/c1lc20606k
    1. Castilho, L. R. (2015). Continuous animal cell perfusion processes: The first step toward integrated continuous biomanufacturing. In G. Subramanian (Ed.), Continuous Processing in Pharmaceutical Manufacturing (pp. 115-154). Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA. https://doi.org/10.1002/9783527673681.ch06
    1. Castilho, L. R., & Medronho, R. A. (2002). Cell retention devices for suspended-cell perfusion cultures. Advances in Biochemical Engineering/Biotechnology, 74, 129-169. https://doi.org/10.1007/3-540-45736-4_7
    1. Castilho, L. R., & Medronho, R. A. (2008). Animal cell separation. In L. R. Castilho, A. M. Moraes, E. F. P. Augusto & M. Butler (Eds.), Animal Cell Technology: From Biopharmaceuticals to Gene Therapy (1st ed.). Nova York: Taylor & Francis.

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