Table 1 Allegro XRS 20 fed batch process conditions.

Cell Line:

Industrial suspension CHO producing mAb biosimilar

Seed:

0.3 - 0.5 × 10⁶ viable cells/mL in 5 L CDFortiCHO (Life Technologies)

Expansion:

Expand at 2 - 4 viable cells/mL to 14 L

Feeding strategy:

CD EfficientFeed C (EFC, Life Technologies), initial feed at 8.0 ± 2.0 M viable cells.mL

Additional bolus feeds every 24 hours for a total of 5 feeds (1.2 L/feed)

Carbon source:

Glucose kept above >2 f/L (bolus target >5f/L)

Antifoam C:

Added as needed

Aeration rate:

1.0 L/min

DO set point:

O₂ control at 40%

pH set point:

7.2 ± 0.05

Agitation:

5 L: 25 RPM, 5°X, 5°Y; 14 - 20: 30 or 35 RPM, 15°X, 5°Y

Temperature:

37°C