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\begin{document}

\title{Real-Time Monitoring of Early Cell Death in Mammalian Cell Culture using
Capacitance Spectroscopy}



\author[1]{Suyang Wu}%
\author[2]{Stephanie Ketcham}%
\author[2]{Claudia Corredor}%
\author[2]{Douglas Both}%
\author[1]{James Drennen}%
\author[1]{Carl Anderson}%
\affil[1]{Duquesne University}%
\affil[2]{Bristol-Myers Squibb Co}%


\vspace{-1em}



  
  \date{May 25, 2024}


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\begin{abstract}
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. 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 that had similar
spectra shape across all probes was first constructed, which improved
the prediction accuracy. Moreover, the global calibration method
included the variance of different probes and scales into the model,
reducing the prediction bias. External parameter orthogonalization also
mitigated the interference from feeding, which further improved the
model performance. 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. In addition, incorporation
of at-line spectra following orthogonal design into the calibration set
is more likely to generate robust calibration models than the
calibration models constructed using the in-line spectra only. This is
advantageous, as at-line spectra collection is easier, faster, and more
material-sparing than in-line spectra collection. The root-mean-square
error of prediction of the final model was 6.56\% (8.42\% of the
prediction range) with an R2 of 92.4\%.%
\end{abstract}%



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\textbf{Hosted file}

\verb`In Line Measurement_V3.docx`  available at \url{https://authorea.com/users/481692/articles/568594-real-time-monitoring-of-early-cell-death-in-mammalian-cell-culture-using-capacitance-spectroscopy}

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