An early stage fed-batch cell culture process we developed was shown to exhibit higher oxygen demand in bioreactor than typically observed in our platform processes. The high oxygen demand is met by increasing agitation and sparging of air and/or oxygen in bioreactors. High sparging rates lead to higher gas entrance velocities (GEV), which in turn can cause cell damage.  This case study demonstrates how the cell culture performance and antibody production were impacted at manufacturing scale by high gas entrance velocities. Further exacerbating this issue, sparger hole plugging at the production scale was observed leading to theoretical GEV of as high as 300 m/s. We investigated gas entrance velocities between 1 and 300 m/s at 2L scale and  examined the impact on cell density, cell viability, and LDH as a surrogate for cell death.  In addition, we also analyzed the cell state under high GEV using flow cytometer to evaluate apoptosis and cell cycle of the cell culture. Extensive trouble shooting studies carried out at small scale confirmed the impact of high GEV on this antibody production process and also pointed to the need for re-design of sparger at manufacturing scale.<br/>

Impact of gas entrance velocity on cell culture performance at small scale and manufacturing scale bioreactors: A case study

253rd National Meeting (2017)

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