Merck

As the biopharmaceutical industry adopts a more strategic view toward manufacturing, there is an increased need for process intensification via unit operation optimization as well as linked and continuous processing. The Life Science business of Merck KGaA, Darmstadt Germany has developed new products and services that meet these challenges and can be implemented into robust manufacturing processes. We offer upstream process intensification solutions to overcome the limitations historically associated with perfusion bioreactors – including low product concentrations, high media costs and overly complex systems. For example, the CHOZN® host cell line platform is leveraged to develop high producing cells capable of stable expression out to more than 90 generations, a requirement for 60- and 90-day perfusion applications¹. In addition, cell culture media is being specifically designed to support high cell density perfusion processes² and our Mobius® single-use bioreactor platform offers the mixing and aeration capabilities to support perfusion processes³. Finally, the Prostak™ family of TFF microfilters have been shown to be effective cell retention devices for such perfusion applications⁴. We are in the final stages of commercializing a unique sterile connector, Lynx® CDR, which allows for multiple sterile connections/disconnections and will facilitate closed processing. Downstream process intensification solutions focus on eliminating current bottlenecks and/or significantly improving process economics. For high-titer mAb processes, continuous multi-column chromatography is an attractive solution for eliminating the protein A bottleneck – especially when employing high capacity, incompressible protein A resins such as our ProSep® Ultra Plus and Eshmuno® A Protein A media⁵. We have further shown that using single-pass filtration to pre-concentrate the product prior to loading onto a continuous protein A capture process is particularly effective in further enhancing the productivity and cost-effectiveness of this capture operation⁶. Post-protein A polishing templates typically employ a combination of bind-elute and flow-through chromatography steps to achieve the necessary removal of process- and product-related impurities. We have demonstrated that an appropriately designed flow-through train is a robust polishing solution for improving process economics of existing batch processes and a key enabler of connected and continuous processes⁷. These technologies work orthogonally and synergistically to remove remaining impurities while minimizing the need to make significant solution condition adjustments, thereby simplifying the manufacturing process. Our studies have further shown that this new flow-through template (with prototype products) can simultaneously reduce total polishing cost of goods by more than 20%, reduce buffer and water consumption by more than 50%, decrease processing time by around 40%, and decrease overall footprint by about 30% in support of existing batch processes – with greater benefits observed for continuous operation⁸. 1. Sealover, N.R., Davis, A.M., Brooks, J.K., George, H.J., Kayser, K.J., and Lin, N. “Engineering Chinese Hamster Ovary (CHO) cells for producing recombinant proteins with simple glycoforms by zinc-finger nuclease (ZNF)-mediated gene knockout of mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (Mgat1).” J. Biotechnol (167) 24-32 2013. 2. Lyons, D., “The future of EX-CELL® Advanced^TM portfolio.” ESACT 24th Meeting. Barcelona, Spain (2015). 3. Rank, D., Martin, C.S., Madrid, L., Phillips, M.W. “mAb product consistency achieved in long duration microfiltration-based CHO perfusion process.” Integrated Continuous Biomanufacturing II. Berkeley, CA, USA (2015). 4. Martin, CS., Padilla-Zamudio, J., Rank, D., McInnis, P., Kozlov, M., Reynolds, S., Madrid, L. “Novel small-scale TFF cell retention device for perfusion cell culture systems.” ESACT 24th Meeting. Barcelona, Spain (2015). 5. Gillespie, C., Kozlov, M., Phillips, M., Potty, A.S.R., Skudas, R., Stone, M., Xenopoulos, A., Dupont, A., Jaber, J., Cataldo, W., “Integrating continuous and single-use methods to establish a new downstream processing platform for monoclonal antibodies.” In: Subramanian, G. (Ed.), Continuous Processing in Biopharmaceutical Manufacturing. Wiley-VCH 2015. 6. Beckett, P. Webinar: “Single-Pass Tangential Flow Filtration: A Critical Operation Within the Continuous Bioprocess” (2015), www.merckmillipore.com/webinars 7. Skudas, R., Beckett, P., Webinar: “Flow through Chromatography and Adsorptive Depth Filtration for Continuous Bioprocessing Applications” (2016), www.merckmillipore.com/webinars 8. Xenopoulos, A. “A new, integrated, continuous purification process template for monoclonal antibodies: Process modeling and cost of goods studies” J. Biotechnol (213), 42-53 2015.

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