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Cytotechnology. May 2003; 42(1): 35–45.
PMCID: PMC3449505
Optimization and control of perfusion cultures using a viable cell probe and cell specific perfusion rates
Jason E. Dowd,1,2 Anthea Jubb,3 K. Ezra Kwok,2 and James M. Piretcorresponding author1
1Biotechnology Laboratory, University of British Columbia, Vancouver, BC Canada
2Department of Chemical and Biological, Engineering, University of British Columbia, Vancouver, BC Canada
3Process Development & Manufacturing, INEX Pharmaceuticals Corporation, Burnaby, BC Canada
James M. Piret, Fax: (604) 822 2114), jpiret/at/chml.ubc.ca.
corresponding authorCorresponding author.
Abstract
Consistent perfusion culture production requires reliable cell retention and control of feed rates. An on-line cell probe based on capacitance was used to assay viable biomass concentrations. A constant cell specific perfusion rate controlled medium feed rates with a bioreactor cell concentration of ~5 × 106 cells mL-1. Perfusion feeding was automatically adjusted based on the cell concentration signal from the on-line biomass sensor. Cell specific perfusion rates were varied over a range of 0.05 to 0.4 nL cell-1 day-1. Pseudo-steady-state bioreactor indices (concentrations, cellular rates and yields) were correlated to cell specific perfusion rates investigated to maximize recombinant protein production from a Chinese hamster ovary cell line. The tissue-type plasminogen activator concentration was maximized (~40 mg L-1) at 0.2 nL cell-1 day-1. The volumetric protein productivity (~60 mg L-1 day-1 was maximized above 0.3 nL cell-1 day-1. The use of cell specific perfusion rates provided a straightforward basis for controlling, modeling and optimizing perfusion cultures.
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Selected References
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