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Nutrient Regulation by Continuous Feeding for Large-scale Expansion of Mammalian Cells in Spheroids
Published on: September 25, 2016
Parallel substrate feeding and pH-control in shaking-flasks
D Weuster-Botz1, J Altenbach-Rehm, M Arnold
1Institute of Biochemical Engineering, Munich University of Technology, Boltzmannstr. 15, 85748, Garching, Germany
Abstract:
An intermittent feeding system for shaking-flasks was developed to close the gap between batch operated shaking-flasks and fed-batch operated as well as pH-controlled stirred tank reactors. A precise syringe pump was connected via a substrate distribution system to individual 2/2-way miniature valves, one for each of up to 16 shaking-flask. The shaking-flasks were equipped with pH-probes. A process computer controls the intermittent feeding of substrates by tracking predefined individual feeding profiles as well as the base (or acid) addition for individual pH-control of the shaking-flasks. Higher concentrations of aerobic cells with higher cellular activities were achieved in fed-batch operated and pH-controlled shaking-flasks as compared to the conventional batch operation. Physiological effects of an intermittent feeding were studied in a stirred tank reactor with a recombinant E. coli strain, which expressed the GDP-mannose-pyrophosphorylase enzyme under the control of the lac-promoter.
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