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New approaches in process monitoring during industrial fermentations are not only limited to classical pH, dO₂ and offgas analysis, but use different in situ and online sensors based on different physical principles to determine biomass, product quality, lysis and far more. One of the very important approaches is the in situ accessibility of viable cell concentration (VCC). This knowledge provides increased efficiency in monitoring and controlling strategies during cultivations. Electrochemical impedance spectroscopy-EIS-is used to monitor biomass in a fermentation of E. coli BL21(DE3), producing a recombinant protein using a fed batch-based approach. Increases in the double layer capacitance (C dl ), determined at frequencies below 1 kHz, are proportional to the increase of biomass in the batch and fed batch phase, monitored in offline and online modes for different cultivations. A good correlation of C dl with cell density is found and in order to get an appropriate verification of this method, different state-of-the-art biomass measurements are performed and compared. Since measurements in this frequency range are largely determined by the double layer region between the electrode and media, rather minor interferences with process parameters (aeration, stirring) are to be expected. It is shown that impedance spectroscopy at low frequencies is a powerful tool for cultivation monitoring.

A Novel Application for Low Frequency Electrochemical Impedance Spectroscopy as an Online Process Monitoring Tool for Viable Cell Concentrations