Understanding and modeling retention of mammalian cells in fluidized bed centrifuges.
Within the last decade, fully disposable centrifuge technologies, fluidized-bed centrifuges (FBC), have been introduced to the biologics industry. The FBC has found a niche in cell therapy where it is used to collect, concentrate, and then wash mammalian cell product while continuously discarding centrate. The goal of this research was to determine optimum FBC conditions for recovery of live cells, and to develop a mathematical model that can assist with process scaleup. Cell losses can occur during bed formation via flow channels within the bed. Experimental results with the kSep400 centrifuge indicate that, for a given volume processed: the bed height (a bed compactness indicator) is affected by RPM and flowrate, and dead cells are selectively removed during operation. To explain these results, two modeling approaches were used: (i) equating the centrifugal and inertial forces on the cells (i.e., a force balance model or FBM) and (ii) a two-phase computational fluid dynamics (CFD) model to predict liquid flow patterns and cell retention in the bowl. Both models predicted bed height vs. time reasonably well, though the CFD model proved more accurate. The flow patterns predicted by CFD indicate a Coriolis-driven flow that enhances uniformity of cells in the bed and may lead to cell losses in the outflow over time. The CFD-predicted loss of viable cells and selective removal of the dead cells generally agreed with experimental trends, but did over-predict dead cell loss by up to 3-fold for some of the conditions. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1520-1530, 2016.
Dept. of Chemical Engineering, Villanova University, Villanova, PA.
William
W
Kelly
Cell Technology Pharmaceutical Development and Manufacturing Sciences, Janssen R&D, Spring House, PA.
Jonathan
J
Rubin
Dept. of Chemical Engineering, Villanova University, Villanova, PA.
Jennifer
J
Scully
Cell Technology Pharmaceutical Development and Manufacturing Sciences, Janssen R&D, Spring House, PA.
Hari
H
Kamaraju
Janssen Infectious Diseases and Vaccines, Leiden, 2333, CN, the Netherlands.
Piotr
P
Wnukowski
Cell Technology Pharmaceutical Development and Manufacturing Sciences, Janssen R&D, Spring House, PA.
Ravinder
R
Bhatia
D019149:Bioreactors; D002460:Cell Line; D002469:Cell Separation; D002498:Centrifugation; D006801:Humans
centrifuge; computational fluid dynamics; counter-flow elutriation; fluidize-bed; fluidized bed centrifuge; kSep; mammalian cells; single-use