Capacitance spectroscopy enables real-time monitoring of early cell death in mammalian cell culture.

Autor: Wu S; Duquesne Center for Pharmaceutical Technology, Duquesne University, Pittsburgh, Pennsylvania, USA.; Duquesne University Graduate School for Pharmaceutical Sciences, Pittsburgh, Pennsylvania, USA., Ketcham SA; Manufacturing Science and Technology, Bristol-Myers Squibb, Devens, Massachusetts, USA., Corredor CC; Pharmaceutical Development, Bristol-Myers Squibb, New Brunswick, New Jersey, USA., Both D; Pharmaceutical Development, Bristol-Myers Squibb, New Brunswick, New Jersey, USA., Drennen JK; Duquesne Center for Pharmaceutical Technology, Duquesne University, Pittsburgh, Pennsylvania, USA.; Duquesne University Graduate School for Pharmaceutical Sciences, Pittsburgh, Pennsylvania, USA., Anderson CA; Duquesne Center for Pharmaceutical Technology, Duquesne University, Pittsburgh, Pennsylvania, USA.; Duquesne University Graduate School for Pharmaceutical Sciences, Pittsburgh, Pennsylvania, USA.
Jazyk: angličtina
Zdroj: Biotechnology journal [Biotechnol J] 2023 Mar; Vol. 18 (3), pp. e2200231. Date of Electronic Publication: 2022 Dec 22.
DOI: 10.1002/biot.202200231
Abstrakt: Background/aims: Previous work developed a quantitative model using capacitance spectroscopy in an at-line setup to predict the dying cell percentage measured from a flow cytometer. This work aimed to transfer the at-line model to monitor lab-scale bioreactors in real-time, waiving the need for frequent sampling and enabling precise controls.
Methods and Results: Due to the difference between the at-line and in-line capacitance probes, direct application of the at-line model resulted in poor accuracy and high prediction bias. A new model with a variable range and offering similar spectral shape across all probes was first constructed, improving prediction accuracy. Moreover, the global calibration method included the variance of different probes and scales in the model, reducing prediction bias. External parameter orthogonalization, a preprocessing method, also mitigated the interference from feeding, which further improved model performance. The root-mean-square error of prediction of the final model was 6.56% (8.42% of the prediction range) with an R 2 of 92.4%.
Conclusion: The culture evolution trajectory predicted by the in-line model captured the cell death and alarmed cell death onset earlier than the trypan blue exclusion test. Additionally, the incorporation of at-line spectra following orthogonal design into the calibration set was shown to generate calibration models that are more robust than the calibration models constructed using the in-line spectra only. This is advantageous, as at-line spectral collection is easier, faster, and more material-sparing than in-line spectra collection.
(© 2022 Wiley-VCH GmbH.)
Databáze: MEDLINE
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