High cell density perfusion process was developed for a cell line with low specific productivity in fed batch process. This work highlights perfusion media development strategy that was successfully developed and optimized based on an existing media for a different cell line. Components hypothesized to be unnecessary were removed to allow for increasing media depth by greater than 2-fold while staying around physiologically relevant media osmolality of 350 mOsm/kg. Mock perfusion studies were used to optimize the concentration of some components such as Ferric Ammonium Citrate, Sodium bicarbonate and amino acids such as glutamine to improve productivity. Additional supplements were identified to improve specific productivity by at least 1.5 to 2-fold although these negatively impacting cell growth. ambr®250 high throughput perfusion system was used to evaluate a biphasic perfusion process using a perfusion media that can support optimal cell growth to reach target cell density followed by a second phase perfusion media that improved specific productivity. Media concentrate strategy was optimized to support optimal performance upon scale up. This perfusion process was then successfully scaled up to a 50 L single-use bioreactor demonstrating similar performance as a 2L bioreactor scale.

Perfusion media development strategies for high intensity perfusion cultures

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.