High cell density is an important factor in achieving high bioreactor productivity. To meet the oxygen demand with density at &gt;100 × 106 cells/mL, a frit sparger is often used. In this presentation, the impact of Pluronic® F68 on a perfusion process using a frit sparger will be discussed. The perfusion process was developed using an alternating tangential flow device with a 0.2 µm Polyethersulfone (PES) hollow fiber filter. Pluronic® F68 at 2 g/L was sufficient in preventing cell damage at gas flow rate of ~0.20 vvm from a drilled hole sparger (0.5 mm) but inadequate at ~0.025 vvm from a frit sparger (20 µm). We demonstrated that increasing Pluronic® F68 concentration to 5 g/L prevented cell death at up to ~0.10 vvm from the frit sparger and was able to maintain high cell density at high viability in the range of 60 – 80 × 106 cells/mL. Such positive effect was demonstrated in both 3- and 200-L bioreactors. Supplementing additional Pluronic® F68 was also effective in restoring cell growth/viability from low viability cultures. We will also discuss the impact of increased Pluronic® F68 concentration on target antibody, host cell proteins, and Pluronic® F68 transmissions across the hollow fiber filter.

Impact of Pluronic® F68 on hollow fiber filter-based perfusion culture performance

255th National Meeting (2018)

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