Mammalian cell culture scale-up and fed-batch control using automated flow cytometry

Area of Science:

• Biotechnology
• Cell Biology
• Process Engineering

Background:

• Mammalian cell culture kinetics are crucial for industrial bioreactor fed-batch strategies.
• Predicting the end of exponential proliferation is a critical parameter for culture scale-up.

Purpose of the Study:

• To demonstrate automated flow cytometry's ability to predict stationary phase in CHO cell cultures.
• To enable automated culture scale-up and improve fed-batch control.

Main Methods:

• Utilized automated flow cytometry to monitor Chinese Hamster Ovary (CHO) cell cultures.
• Tracked the increase in the non-viable sub-population as an indicator for stationary phase onset.

Main Results:

• An increase in the non-viable cell sub-population predicted stationary phase approximately 40 hours in advance.
• Automated scale-up resulted in significantly higher total cell counts compared to manual scale-up.
• Observed significant variation in proliferation rates across individual culture expansions.

Conclusions:

• Automated flow cytometry provides critical process parameters for new control strategies in cell culture.
• This approach supports the FDA's Process Analytical Technology (PAT) initiative for enhanced product quality.
• Enables reliable, reproducible control of fed-batch additions and culture expansion.