Journal
CYTOTECHNOLOGY
Volume 68, Issue 3, Pages 399-408Publisher
SPRINGER
DOI: 10.1007/s10616-014-9791-3
Keywords
Flow cytometry; Online; At-line; Automation; CHO cells; PAT; Monitoring; Apoptosis
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Funding
- Enterprise Ireland for the apPAT project
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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.
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