Development of an N-1 perfusion medium to intensify seed train operation
October 6-10, 2019
Seed train expansion of cells before the final production step is often time-consuming and a major source of process variability. For the intensification of seed train operations there are several opportunities discussed across the biopharma industry today. One of the possibilities is to operate N-1 bioreactors in perfusion mode to shorten timelines and improve bioreactor utilization. In this work, we investigated the influence of using an expansion medium especially designed for N-1 perfusion to gain optimal results in the whole manufacturing campaign.
For screening and designing an N-1 perfusion expansion medium, a scale down model which represents a typical production campaign, including the seed train, was established. Expansion medium prototypes were combined with different production media in the final production step, e.g. EX-CELL® Advanced HD Perfusion as medium designed for high-density perfusion, and Cellvento® 4CHO Medium and 4FEED as exemplary fed-batch process.
After determining a suitable expansion medium formulation, the prototype was evaluated for solubility and streamlined to ensure a simple hydration and robust supply chain. Afterwards, results were confirmed using a simulated manufacturing process using benchtop bioreactor systems, showing that the positive impact of the expansion medium on the final yield is present both when using perfusion or fed-batch as final production stage.
Our results indicate that using the right companion medium in seed train expansion - specifically designed for the purpose - can prepare the cells optimally for the final N-stage and increase productivity while using low CSPRs. Combining these findings with the application of a perfused N-1 step in the manufacturing campaign leads to a great opportunity for the intensification of the whole upstream process.
Luis Fernando Ayala Solares, Caroline Ströder, Christian Schultheiss, Jana Glawion, Melanie Feigenspan, Jochen B. Sieck, and Luis Fernando Ayala Solares, "Development of an N-1 perfusion medium to intensify seed train operation" in "Integrated Continuous Biomanufacturing IV", Veena Warikoo, Roche, USA Alois Jungbauer, BOKU, Austria Jon Coffman, Boehringer Ingelheim, USA Jason Walther, Sanofi, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/biomanufact_iv/6