title: Online flow cytometry for monitoring apoptosis in mammalian cell cultures as an application for process analytical technology.
abstract: Apoptosis is the main driver of cell death in bioreactor suspension cell cultures during the production of biopharmaceuticals from animal cell lines. It is known that apoptosis also has an effect on the quality and quantity of the expressed recombinant protein. This has raised the importance of studying apoptosis for implementing culture optimization strategies. The work here describes a novel approach to obtain near real time data on proportion of viable, early apoptotic, late apoptotic and necrotic cell populations in a suspension CHO culture using automated sample preparation in conjunction with flow cytometry. The resultant online flow cytometry data can track the progression of apoptotic events in culture, aligning with analogous manual methodologies and giving similar results. The obtained near-real time apoptosis data are a significant improvement in monitoring capabilities and can lead to improved control strategies and research data on complex biological systems in bioreactor cultures in both academic and industrial settings focused on process analytical technology applications.
authors.affiliation: School of Chemical and Bioprocess Engineering and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.|Sanford Burnham Medical Research Institute, Protein Production and Analysis Core, 6400 Sanger Road, Medical City, Orlando, FL, 32832, USA.
authors.forename: Darrin
authors.initials: D
authors.lastname: Kuystermans
authors.affiliation: School of Chemical and Bioprocess Engineering and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Mohd
authors.initials: M
authors.lastname: Avesh
authors.affiliation: School of Chemical and Bioprocess Engineering and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland. m.al-rubeai@ucd.ie.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Al-Rubeai
keywords: Apoptosis; At-line; Automation; CHO cells; Flow cytometry; Monitoring; Online; PAT