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adequate liquid homogenization in large scale bioreactors Typical values of energy dissipation rates are two orders below that used in bacterial fermentations One of the very few mixing and aeration studies in large scale bioreactors under animal cell conditions, other than the one reported by Kiss et al (1994), was reported by Nienow et al (1996) and Langheinrich et al (1998) These authors conducted comprehensive studies of homogenization in a commercial scale 8 m3 bioreactor and in a geometrically similar scaled-down 061 m mixing tank The 8 m3 bioreactor was operated in industrial applications for cell culture growth at a maximum speed of 60 rpm with a single Rushton turbine with an unusually low D/T and clearance ratios (C/T) of 022 and an aeration rate via a pipe sparger of 0005 vvm Under such conditions the energy dissipation rates due to agitation and aeration were 11.



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5 10 3 and 2 10 3 W/kg, respectively First, they showed that under such operating conditions the mixing time was >200 s for H/T = 13 and that the mixing time correlations derived in the literature for 1 < H/T ratio < 3 (Cooke et al, 1988) were capable of predicting the experimentally derived mixing times in the 8 m3 bioreactor, even though the energy dissipation and aeration rates were two orders of magnitude lower than those used to derive the correlations Second, they demonstrated that mixing at the top of the bioreactor was poor, due to the inability of the upper recirculation loop to reach the top of the bioreactor, and also that the aerated mixing time was signi cantly shorter than under unaerated conditions at the top of the bioreactor, due to the energy dissipated by bubble disengagement and convective ow.

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Suggested strategies for signi cant improvement in liquid blending arising from experimentation in the scaled-down tank included retro tting the small diameter high power number Rushton with a large diameter low power number axial ow impeller such as an up-pumping Chemineer HE3 or a Prochem Max o T, the use of increased energy dissipation rates, and an increased impeller clearance ratio C/T of 05 For caustic pH control, alkali is generally added at the liquid surface at the later stages of cell culture processes, and it is here that pH excursions are known to occur, especially if liquid blending is poor Langheinrich and Nienow (1999) also used the geometrically scaled down 061 m tank to investigate possible pH excursions in the 8 m3 bioreactor At the operating conditions described earlier, they found signi cant pH excursions of up to 0.

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8 pH unit at the point of alkali addition in a buffered system using both pH probe measurements and ow visualization studies Such excursions can be detrimental for cell viability and product formation (Brown and Birch, 1996; Osman et al, 2001, 2002) The pH variations were absent when alkali was added to the impeller region They also found that an increase in the overall energy dissipation rate did not have a signi cant impact on the pH excursions given the substantial differences in the local energy dissipation rates at the liquid surface and the impeller region The authors recommended that the pH excursions could, realistically, be avoided only with alkali added to the impeller region via a pipe Traditionally, this option has not been implemented in industrial scale bioreactors due to clean in place (CIP) considerations..

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