An automated laboratory‐scale methodology for the generation of sheared mammalian cell culture samples

Abstract

Continuous disk-stack centrifugation is typically used for the removal of cells and cellular debris from mammalian cell culture broths at manufacturing-scale. The use of scale-down methods to characterize disk-stack centrifugation performance enables substantial reductions in material requirements and allows a much wider design space to be tested than is currently possible at pilotscale. The process of scaling down centrifugation has historically been challenging due to the difficulties in mimicking the Energy Dissipation Rates (EDRs) in typical machines. This paper describes an alternative and easy-to-assemble automated capillary-based methodology to generate levels of EDRs consistent with those found in a continuous disk-stack centrifuge. Variations in EDR were achieved through changes in capillary internal diameter and the flow rate of operation through the capillary. The EDRs found to match the levels of shear in the feed zone of a pilot-scale centrifuge using the experimental method developed in this paper (2.4 × 10 5 W/kg) are consistent with those obtained through previously published computational fluid dynamic studies (2.0 × 10 5 W/kg). Furthermore, this methodology can be incorporated into existing scale-down methods to model the process performance of continuous disk-stack centrifuges. This was demonstrated through the characterization of culture hold time, culture temperature and EDRs on centrate quality.

Online control of cell culture redox potential prevents antibody interchain disulfide bond reduction

“…For some processes, mAb reduction is not observed until after harvest. The approach of controlling the cell culture redox potential can be utilized to predict the likelihood of mAb reduction during harvest by incorporating the appropriate harvest model (Joseph et al, 2017). A new correlation can be developed based on the preharvest cell culture redox potential and the percent of intact mAb observed after harvest.…”

Enhanced harvest performance predictability through advanced multivariate data analysis of mammalian cell culture particle size distribution

“…The capillary features a very small inner OD of 0.01′ (0.0254 cm). Further details about the CSD are discussed by Westoby et al (2011) and Joseph et al (2015, 2017).…”

“…The cells can lose membrane integrity towards the end of cell culture as the cell culture viability drops. Cells can also lyse due to mechanical forces such as the shear experienced at the centrifuge feedzone, where the feed is accelerated to the bowl speed (Joseph et al, 2017). Rapid detection of cell lysis at the end of cell culture prior and post centrifugation would help predict and understand the conditions that lead to product degradation and devise informed strategies to minimize quality deviations.…”