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. 2003 May;42(1):35–45. doi: 10.1023/A:1026192228471

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, James M Piret 1,✉
PMCID: PMC3449505  PMID: 19002926

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.

Keywords: Cell specific perfusion feed rates, CHO, Control, Optimization, t-PA, Viable cell probe

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