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Online flow cytometry for monitoring apoptosis in mammalian cell cultures as an application for process analytical technology

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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.

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Acknowledgments

We thank Christian Lavarreda of MSP Corporation for the use of the Flow CytoPrep 5000 unit. This work was supported by funds from Enterprise Ireland for the apPAT project.

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Author notes

  1. Darrin Kuystermans

    Present address: Sanford Burnham Medical Research Institute, Protein Production and Analysis Core, 6400 Sanger Road, Medical City, Orlando, FL, 32832, USA

Authors and Affiliations

  1. School of Chemical and Bioprocess Engineering and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland

    Darrin Kuystermans, Mohd Avesh & Mohamed Al-Rubeai

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  1. Darrin Kuystermans
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  2. Mohd Avesh
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  3. Mohamed Al-Rubeai
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Correspondence to Mohamed Al-Rubeai.

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Kuystermans, D., Avesh, M. & Al-Rubeai, M. Online flow cytometry for monitoring apoptosis in mammalian cell cultures as an application for process analytical technology. Cytotechnology 68, 399–408 (2016). https://doi.org/10.1007/s10616-014-9791-3

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