Cascaded processing enables continuous upstream processing with E. coli BL21(DE3)
Stefan Kittler, Christoph Slouka, Andreas Pell, Roman Lamplot, Mihail Besleaga, Sarah Ablasser, Christoph Herwig, Oliver Spadiut, Julian Kopp
Abstract
In many industrial sectors continuous processing is already the golden standard to maximize productivity. However, when working with living cells, subpopulation formation causes instabilities in long-term cultivations. In cascaded continuous cultivation, biomass formation and recombinant protein expression can be spatially separated. This cultivation mode was found to facilitate stable protein expression using microbial hosts, however mechanistic knowledge of this cultivation strategy is scarce. In this contribution we present a method workflow to reduce workload and accelerate the establishment of stable continuous processes with E. coli BL21(DE3) exclusively based on bioengineering methods.