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Scaling up the HEK293 DKO seed train from a 1 L shake flask to controlled 2 L bioreactors. Bioreactor #1: pH setpoint of 7 with a deadband of ±0.03 and a DO setpoint of 30%. Bioreactor #2: pH setpoint of 7 with a deadband of ±0.4 and a DO setpoint of 60%. Passaging the 1 L shake flask and 2 L bioreactors every 3–4 days for 25 days: (A) VCD and viability, (B) glucose and lactate, (C) offline pH, and (D) pO2. (E) Day 7 transfection titers from 30 mL tubespins.
