Innovative fixed bed bioreactor platform: Enabling linearly scalable adherent cell biomanufacturing with real‐time biomass prediction from nutrient consumption

• Published in: Biotechnology journal 2024-08, Vol.19 (8), p.e2300635-n/a
• Main Authors: Goral, Vasiliy N., Hong, Yulong, Scibek, Jeffery J., Sun, Yujian, Romeo, Lori E., Rao, Abhijit, Manning, Daniel, Zhou, Yue, Schultes, Joel A., Tjong, Vinalia, Pikula, Dragan, Krebs, Kathleen A., Ferrie, Ann M., Kramel, Stefan, Weber, Jennifer L., Upton, Todd M., Fang, Ye, Melkoumian, Zara
• Format: Article
• Language: English
• Subjects: adeno‐associated virus vector; adherent cell biomanufacturing; Animals; ascent; Biomass; Bioreactors; Cell Adhesion; Cell Culture Techniques - instrumentation; Cell Culture Techniques - methods; Cell Proliferation; CHO Cells; computational fluid dynamics; Cricetulus; Equipment Design; fixed bed bioreactor; Glucose - metabolism; Humans; Hydrodynamics; oxygen uptake rate
• ISSN: 1860-6768; 1860-7314; 1860-7314
• DOI: 10.1002/biot.202300635

Scalable single‐use adherent cell‐based biomanufacturing platforms are essential for unlocking the full potential of cell and gene therapies. The primary objective of this study is to design and develop a novel fixed bed bioreactor platform tailored specifically for scaling up adherent cell culture. The bioreactor comprises a packed bed of vertically stacked woven polyethylene terephthalate mesh discs, sandwiched between two‐fluid guide plates. Leveraging computational fluid dynamics modeling, we optimized bioreactor design to achieve uniform flow with minimal shear stress. Residence time distribution measurements demonstrated excellent flow uniformity with plug flow characteristics. Periodic media sampling coupled with offline analysis revealed minimal gradients of crucial metabolites (glucose, glutamine, lactate, and ammonia) across the bioreactor during cell growth. Furthermore, the bioreactor platform demonstrated high performance in automated cell harvesting, with ≈96% efficiency and ≈98% viability. It also exhibited linear scalability in both operational parameters and performance for cell culture and adeno‐associated virus vector production. We developed mathematical models based on oxygen uptake rates to accurately predict cell growth curves and estimate biomass in real‐time. This study demonstrates the effectiveness of the developed fixed‐bed bioreactor platform in enabling scalable adherent cell‐based biomanufacturing with high productivity and process control. Graphical and Lay Summary We designed a novel bioreactor system to improve large‐scale cell production for advanced therapies. It ensures uniform cell growth conditions, efficient harvesting, and easy scalability. This innovation could significantly enhance the manufacturing efficiency and accessibility of cell and gene therapy.