Abstract
As part of the process-characterization campaign of a candidate vaccine product, a recently developed class of three-level designs—definitive-screening designs—was employed to select a quadratic model that describes the effect of six input process parameters, including protein concentration, formaldehyde-to-protein ratio, lysine concentration, reaction duration, pH, and reaction temperature, on a formylation protein-crosslinking reaction. This design requires only 17 experimental runs. The resulting model was then used to simulate 10,000 runs that account for the variability in the inputs expected on manufacturing scale. The extent of protein polymerization was predicted to be within specifications for all simulated runs, demonstrating the robustness of the unit operation for subsequent process validation and future commercial manufacturing.
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Acknowledgments
We thank Marina Eiting, Diana Olschewski, and Lorenz Hasler of Lonza Ltd. as well as Jesse McCool, Stephen Palmieri, Peychii Lee, and Qi Xu of Lonza Biologics Inc. for process and analytical development. We thank Stephen Collins, Friedhelm Helling, Marilena Paludi, Alessandro Bartalesi, Frederica Sponga, and Valeria Onnis of Novartis Vaccines for technical guidance.
Conflict of interest
Process development and validation are core commercial activities of Lonza Inc.
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Erler, A., de Mas, N., Ramsey, P. et al. Efficient biological process characterization by definitive-screening designs: the formaldehyde treatment of a therapeutic protein as a case study. Biotechnol Lett 35, 323–329 (2013). https://doi.org/10.1007/s10529-012-1089-y
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DOI: https://doi.org/10.1007/s10529-012-1089-y