Integrated process development ‐ quality by design compliant evaluation of upstream variations at the microscale level

Abstract

AbstractBACKGROUNDEvaluation of optimized upstream conditions is often solely based on titers that do not reflect the downstream process and, consequently, the final purity and yield. This is especially critical for proteins expressed as inclusion bodies (IBs) because the subsequent downstream process is more complex than for soluble proteins. A miniaturized process development platform representing the entire downstream process at a microscale level combined with a multi‐fermenter system for bioprocess screening enables fast investigation, using quality by design (QbD) measures, of the quality of feedstock compositions that are altered by upstream conditions.RESULTSHigh‐throughput methods were integrated for an entire process chain, including upstream and downstream processing, for production of a recombinant protein as IBs. The miniaturized process chain consisted of cell disruption, IB harvesting, solubilization, refolding, and chromatographic purification. This enables processing and evaluation of 16 different fermentation conditions within 7 days. Only 2 g of initial biomass was required for evaluation of an individual fermentation. This resulted in a 15‐fold reduction of time and a 100‐fold reduction of material compared with common bench scale experiments.CONCLUSIONThis microscale process chain mimics the bench process and is able to differentiate purity variations as low as 2%. © 2017 Society of Chemical Industry