In the biopharmaceutical industry, the use of mammalian cells to produce therapeutic proteins is becoming increasingly widespread. Monitoring of these cultures via different analysis techniques is essential to ensure a good quality product while respecting good manufacturing practice (GMP) regulations. Process Analytical Technologies (PAT) tools provide real-time measurements of the physiological state of the culture and enable process automation. Dielectric spectroscopy is a PAT that can be used to monitor the viable cell concentration (VCC) of living cells after processing raw permittivity data. Several modeling approaches exist and estimate biomass with different accuracy. The accuracy of the Cole-Cole and Maxwell Wagner's equations are studied here in the determination of the VCC and cell radius in Chinese hamster ovary (CHO) culture. A sensitivity analysis performed on the parameters entering the equations highlighted the importance of the cell specific parameters such as internal conductivity (σi ) and membrane capacitance (Cm ) in the accuracy of the estimation of VCC and cell radius. The most accurate optimization method found to improve the accuracy involves in-process adjustments of Cm and σi in the model equations with samplings from the bioreactor. This combination of offline and in situ data improved the estimation precision of the VCC by 69% compared to a purely mechanistic model without offline adjustments.
Keywords: Cole-Cole model; cell culture; dielectric spectroscopy; in situ sensors; real-time monitoring.
© 2023 The Authors. Biotechnology Journal published by Wiley-VCH GmbH.