title: 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 characterise 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 pilot-scale. 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 (2.4×105W/Kg) are consistent with those obtained through previously published computational fluid dynamic (CFD) studies (2.0×105W/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 characterisation of culture hold time, culture temperature and EDRs on centrate quality.
authors.affiliation: The Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, London, UK.
authors.forename: Adrian
authors.initials: A
authors.lastname: Joseph
authors.affiliation: The Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, London, UK.
authors.forename: Stephen
authors.initials: S
authors.lastname: Goldrick
authors.affiliation: MedImmune, Gaithersburg Headquarters, Gaithersburg, MD, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Mollet
authors.affiliation: MedImmune, Milstein Building, Granta Park, Cambridge, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Turner
authors.affiliation: MedImmune, Gaithersburg Headquarters, Gaithersburg, MD, USA.
authors.forename: Jean
authors.initials: J
authors.lastname: Bender
authors.affiliation: MedImmune, Milstein Building, Granta Park, Cambridge, UK.
authors.forename: David
authors.initials: D
authors.lastname: Gruber
authors.affiliation: The Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, London, UK.
authors.forename: Suzanne S
authors.initials: SS
authors.lastname: Farid
authors.affiliation: The Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, London, UK.
authors.forename: Nigel
authors.initials: N
authors.lastname: Titchener-Hooker
mesh_terms: D000818:Animals; D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods*; D001696:Biomechanical Phenomena; D001709:Biotechnology; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002498:Centrifugation; D006224:Cricetinae; D003412:Cricetulus; D004867:Equipment Design
keywords: Capillary shear; Disk-stack centrifuge; Energy dissipation rate; Mammalian cell; Scale-down