title: Establishment of a fully automated microtiter plate-based system for suspension cell culture and its application for enhanced process optimization.
abstract: We developed an automated microtiter plate (MTP)-based system for suspension cell culture to meet the increased demands for miniaturized high throughput applications in biopharmaceutical process development. The generic system is based on off-the-shelf commercial laboratory automation equipment and is able to utilize MTPs of different configurations (6-24 wells per plate) in orbital shaken mode. The shaking conditions were optimized by Computational Fluid Dynamics simulations. The fully automated system handles plate transport, seeding and feeding of cells, daily sampling, and preparation of analytical assays. The integration of all required analytical instrumentation into the system enables a hands-off operation which prevents bottlenecks in sample processing. The modular set-up makes the system flexible and adaptable for a continuous extension of analytical parameters and add-on components. The system proved suitable as screening tool for process development by verifying the comparability of results for the MTP-based system and bioreactors regarding profiles of viable cell density, lactate, and product concentration of CHO cell lines. These studies confirmed that 6 well MTPs as well as 24 deepwell MTPs were predictive for a scale up to a 1000 L stirred tank reactor (scale factor 1:200,000). Applying the established cell culture system for automated media blend screening in late stage development, a 22% increase in product yield was achieved in comparison to the reference process. The predicted product increase was subsequently confirmed in 2 L bioreactors. Thus, we demonstrated the feasibility of the automated MTP-based cell culture system for enhanced screening and optimization applications in process development and identified further application areas such as process robustness. The system offers a great potential to accelerate time-to-market for new biopharmaceuticals. Biotechnol. Bioeng. 2017;114: 113-121. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Roche Diagnostics GmbH-Pharmaceutical Biotech Production and Development, Nonnenwald 2, 82377 Penzberg, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Markert
authors.affiliation: Roche Diagnostics GmbH-Pharmaceutical Biotech Production and Development, Nonnenwald 2, 82377 Penzberg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Joeris
mesh_terms: D000818:Animals; D001331:Automation / Q000295:instrumentation*; D061249:Batch Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004867:Equipment Design; D005240:Feasibility Studies
keywords: biopharmaceutical drug development; fed-batch CHO suspension cultures; high-throughput process optimization; industrial cell culture automation; scale up prediction; shaken microtiter plate