Lysis-free separation of hybridoma cells by continuous disc stack centrifugation.

Cytotechnology
H TebbeJ Lehmann

Abstract

The non-destructive removal of hybridoma cells from fermentation broth with an improved disc stack centrifuge (CSA1, Westfalia Separator AG, Oelde, Germany) was investigated. The centrifuge was equipped with a hydrohermetic feed system, which allowed a gentle, shearless acceleration of the cells inside the bowl. No significant cell damage was observed during the separation of hybridoma cells from repeated batch fermentation in 100 liter scale. In the clarified liquid phase there was no increase in Lactate-Dehydrogenase (LDH) activity. Consequently, there was no increased exposure of the product to intracellular components.Due to continuous operation with a periodic and automatic discharge of sediment, a high throughput was achieved without any considerable loss of product. The clarification for mammalian cells was in the range of 99% to 99.9%, depending on the operating conditions. The content of cell debris and other small particles decreased about 30 to 50%, depending on the particle load in the feed stream. The centrifuge was fully contained; cleaning and sterilizing in place possible. Therefore, the decice could be integrated easily into the fermentation process.

References

Jan 1, 1991·Cytotechnology·H BüntemeyerJ Lehmann
Jan 1, 1994·Cytotechnology·W Berthold, R Kempken
Jan 1, 1994·Cytotechnology·F GudermannJ Lehmann
Jan 1, 1995·Journal of Industrial Microbiology·R KempkenW Berthold
Apr 20, 1995·Biotechnology and Bioengineering·R KempkenW Berthold

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Citations

Sep 4, 2015·Biotechnology and Bioengineering·Anna SenczukYinges Yigzaw
Jul 29, 2004·Biotechnology and Bioengineering·E N Lightfoot, J S Moscariello
Apr 18, 2020·Biotechnology Journal·Jure SencarAlois Jungbauer

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