Evaluation of an n-1 perfusion/high-seed fed-batch technology for the intensification of recombinant protein production processes: From AMBR250 to 200 L STR
May 6-11, 2018
Antibody production processes can be accelerated by considerably increasing the starting cell concentration in the production fed-batch bioreactor (high-seed FB). Combining a perfusion bioreactor at the n-1 stage, FB production bioreactors with initial cell concentrations of 10 × 106 cell/mL can be achieved. The resulting time-shortening at the production bioreactor (n stage) can potentially generate a 30 % increase in manufacturing capacity while yielding comparable product quality . While this has been proven in 5 L benchtop bioreactors, a platform covering a broader range of scales can facilitate the process intensification activities from early stage process development up to manufacturing. In this study, the production of a recombinant protein using a high-seed FB strategy was assessed in AMBR250, 2 L and 200 L bioreactor scales. An alternating tangential flow (ATF) perfusion system was used to expand the producer cells to target concentrations ≥ 60 × 106 cells/mL in the corresponding n-1 bioreactors. Production FB bioreactors were then inoculated with 10 × 106 cells/mL and operated up to 9 to 12 days using an automated feeding profile based on either at-line or on-line bio-volume measurements. Optimization of the feeding profile and rate led to comparable antibody productivities and quality throughout the different scales and with respect to the conventional 14-day FB process. Overall, the application of a high-seed FB strategy allowed for a reduction of the processing time and proved the intended increase in the overall production capacity. Also, the assessment of the high-seed FB strategy throughout different scales set the base for the establishment of a robust technology that fulfills the requirements of different products during process intensification.  Yang WC, Lu J, Kwiatkowski C, Yuan H, Kshirsagar R, Ryll T, et al. Perfusion seed cultures improve biopharmaceutical fed-batch production capacity and product quality. Biotechnol Prog. 2014;30:616-25.
Daniel Vazquez Ramirez, Marc Hein, Norbert Schulze, Yali Zhang, and Oliver Krämer, "Evaluation of an n-1 perfusion/high-seed fed-batch technology for the intensification of recombinant protein production processes: From AMBR250 to 200 L STR" in "Cell Culture Engineering XVI", A. Robinson, PhD, Tulane University R. Venkat, PhD, MedImmune E. Schaefer, ScD, J&J Janssen Eds, ECI Symposium Series, (2018). https://dc.engconfintl.org/ccexvi/70