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

High cell density is an important factor in achieving high bioreactor productivity. To meet the oxygen demand with density at >100 × 10 cells/mL, a frit sparger is often used. In this study, the impact of Pluronic F68 on a perfusion process using a frit sparger was studied. The perfusion process was developed using an alternating tangential flow device with a 0.2 µm 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). Increase of 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 × 10 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. Increased Pluronic F68 concentration had no adverse impact on target antibody, HCP, and Pluronic F68 transmissions.