Integrated continuous production of recombinant therapeutic proteins

Abstract

In the current environment of diverse product pipelines, rapidly fluctuating market demands and growing competition from biosimilars, biotechnology companies are increasingly driven to develop innovative solutions for highly flexible and cost-effective manufacturing. To address these challenging demands, integrated continuous processing, comprised of high-density perfusion cell culture and a directly coupled continuous capture step, can be used as a universal biomanufacturing platform. This study reports the first successful demonstration of the integration of a perfusion bioreactor and a four-column periodic counter-current chromatography (PCC) system for the continuous capture of candidate protein therapeutics. Two examples are presented: (1) a monoclonal antibody (model of a stable protein) and (2) a recombinant human enzyme (model of a highly complex, less stable protein). In both cases, high-density perfusion CHO cell cultures were operated at a quasi-steady state of 50-60 × 10(6) cells/mL for more than 60 days, achieving volumetric productivities much higher than current perfusion or fed-batch processes. The directly integrated and automated PCC system ran uninterrupted for 30 days without indications of time-based performance decline. The product quality observed for the continuous capture process was comparable to that for a batch-column operation. Furthermore, the integration of perfusion cell culture and PCC led to a dramatic decrease in the equipment footprint and elimination of several non-value-added unit operations, such as clarification and intermediate hold steps. These findings demonstrate the potential of integrated continuous bioprocessing as a universal platform for the manufacture of various kinds of therapeutic proteins.

Perfusion Processes

“…These processes are more flexible and require lower investment costs compared to the large bioreactor facilities. Processes integrating continuous up-stream and down-stream operations are explored and will probably be the processes of tomorrow (Grandics et al 1991;Kumar et al 2006;Warikoo et al 2012;Vogel et al 2012).…”

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

“…In most cases, to achieve and maintain cell density at [30 9 10 6 cells/mL, medium exchange (i.e., perfusion) is required for nutrients replenishment and by-products removal. Perfusion cultures could be operated at different cell densities (ranging from 10-20 to [100 9 10 6 cells/mL) with different perfusion rates and cell bleed rates, and their productivity has been incrementally improved in recent years, thanks to the advancements in cell line, media, and perfusion technology [9][10][11]. Recently, another operational mode, concentrated fed-batch (CFB), has been re-introduced as an effective approach to significantly increase product titer in the bioreactor [12][13][14].…”

IMAC capture of recombinant protein from unclarified mammalian cell feed streams

“…As yield is a function of the number of processes (Farid, 2011), it is advantageous to remove process steps where possible. Process efficiency can be achieved by improved integration of steps immediately following fermentation (Warikoo et al, 2012), with the added benefit of reducing process volumes, CAPEX, operator costs, and consumables.…”