Abstract
Objective
to develop a new strategy combining near-infrared (NIR) and dielectric spectroscopies for real-time monitoring and in-depth characterizing populations of Chinese hamster ovary cells throughout cultures performed in bioreactors.
Results
Spectral data processing was based on off-line analyses of the cells, including trypan blue exclusion method, and lactate dehydrogenase activity (LDH). Viable cell density showed a linear correlation with permittivity up to 6 × 106 cells ml−1, while a logarithmic correlation was found between non-lysed dead cell density and conductivity up to 107 cells ml−1. Additionally, partial least square technique was used to develop a calibration model of the supernatant LDH activity based on online NIR spectra with a RMSEC of 55 U l−1. Considering the LDH content of viable cells measured to be 110 U per 109 cells, the lysed dead cell density could be then estimated. These calibration models provided real-time prediction accuracy (R2 ≥ 0.95) for the three types of cell populations.
Conclusion
The high potential of a dual spectroscopy strategy to enhance the online bioprocesses characterization is demonstrated since it allows the simultaneous determination of viable, dead and lysed cell populations in real time.
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Acknowledgments
This research has been supported by the French National Research Agency (SPECTRE Project) for Franck Courtès post-doctoral Grant and experimental costs.
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Courtès, F., Ebel, B., Guédon, E. et al. A dual near-infrared and dielectric spectroscopies strategy to monitor populations of Chinese hamster ovary cells in bioreactor. Biotechnol Lett 38, 745–750 (2016). https://doi.org/10.1007/s10529-016-2036-0
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DOI: https://doi.org/10.1007/s10529-016-2036-0