Optimization of an acoustic cell filter with a novel air-backflush system

Area of Science:

• Biotechnology
• Bioprocessing Engineering

Background:

• Perfusion processes are crucial for increasing mammalian cell production capacity.
• Cell retention devices are essential components of perfusion systems.
• Traditional cell retention methods can cause cell damage due to pumping and suboptimal conditions.

Purpose of the Study:

• To develop and evaluate a novel air-backflush operating mode for acoustic cell separators.
• To eliminate cell pumping and reduce potential cell damage in perfusion cultures.
• To define reliable operating ranges and optimize performance for acoustic cell separation.

Main Methods:

• A novel air-backflush mode was implemented in a 10L acoustic cell separator.
• Systematic testing was conducted at cell densities of 10(7) cells/mL.
• Separation performance was evaluated across various airflow rates, backflush frequencies, power settings, and duty cycles.

Main Results:

• Consistent separation performance was achieved with airflow rates from 0-15 L/min and backflush frequencies of 10-40 h(-1).
• Optimized conditions (8W power, 4.5s stop, 45s run) at 10 L/day perfusion rate yielded 92 +/- 0.3% separation efficiency.
• The system was successfully applied in a 110-day CHO cell perfusion culture with 95% viability.

Conclusions:

• The air-backflush mode effectively prevents cell damage by eliminating the need for pumping.
• This technology offers reliable and efficient cell retention for perfusion bioprocesses.
• The developed method supports long-term, high-viability mammalian cell cultures.