Multivariate data analysis of capacitance frequency scanning for online monitoring of viable cell concentrations in small-scale bioreactors.

Viable cell concentration (VCC) is one of the most important process attributes during mammalian cell cultivations. Current state-of-the-art measurements of VCC comprise offline methods which do not allow for continuous process data. According to the FDA's process analytical technology initiative, process monitoring and control should be applied to gain process understanding and to ensure high product quality. In this work, the use of an inline capacitance probe to monitor online VCCs of a mammalian CHO cell culture process in small-scale bioreactors (250 mL) was investigated. Capacitance sensors using single frequency are increasingly common for biomass monitoring. However, the single-frequency signal corresponds to the cell polarization that represents the viable cell volume. Therefore single-frequency measurements are dependent on cell diameter changes. Measuring the capacitance across various frequencies (frequency scanning) can provide information about the VCC and cope with changing cell diameter. Applying multivariate data analysis on the frequency scanning data successfully enabled direct online monitoring of VCCs in this study. The multivariate model was trained with data from 5 standard cultivations. The model provided a prediction of VCCs with relative errors from 5.5 to 11%, which is a good agreement with the acceptance criterion based on the offline reference method accuracy (approximately 10% relative error) and strongly improved compared with single-frequency results (16 to 23% relative error). Furthermore, robustness trials were conducted to demonstrate the model's predictive ability under challenging conditions. The process deviations in regard to dilution steps and feed variations were detected immediately in the online prediction of the VCC with relative errors between 6.7 and 13.2%. Thus in summary, the presented method on capacitance frequency scanning demonstrates its suitability for process monitoring and control that can save batches, time, and cost. Graphical abstract.

Capacitance sensors in cell-based bioprocesses: online monitoring of biomass and more.

Biocapacitance measurement has emerged as a widely used technique for monitoring bioprocesses that involve living cells. Over time, hardware and software developments have enabled this method to move from food towards biopharma industries for improved characterisation, monitoring and control of the bioprocess, even in strictly regulated production environments. In alignment with the general trends in biopharma towards new modalities such as virus-based and cell-based therapies, biocapacitance measurement is entering this area and provides new opportunities for process development and control. Based on the recent progress, the authors strongly believe that even though biocapacitance measurement is a mature, established technology for online biomass monitoring, the nearest future will bring its new and exciting developments and applications that will enhance bioprocess understanding and bring new solutions for enhanced process understanding, monitoring and control.