Development and characterisation of a new fluorescence sensor for online monitoring of bioprocesses

Abstract

Fluorescence spectroscopy is a highly sensitive and non-invasive technique for the identification of characteristic process states and for the online monitoring of substrate and product concentrations. Nevertheless, fluorescence sensors are mainly used in academic studies and are not well implemented for monitoring of industrial production processes. In this work, we present a newly developed robust online fluorescence sensor that facilitates the analysis of fluorescence measurements. The set-up of the sensor was miniaturised and realised without any moveable part to be robust enough for application in technical environments. It was constructed to measure only the three most important biologic fluorophores (tryptophan, NADH and FAD/FMN), resulting in a significant data reduction compared to conventional a 2-D fluorescence spectrometer. The sensor performance was evaluated by calibration curves and selectivity tests. The measuring ranges were determined as 0.5-50 μmolL-1 for NADH and 0.0025-7.5 μmolL-1 for BSA and riboflavin. Online monitoring of batch cultivations of wild-type Escherichia coli K1 in a 10 L bioreactor scale were performed. The data sets were analysed using principal component analysis and partial least square regression. The recorded fluorescence data were successfully used to predict the biomass of an independent cultivation (RMSEP 4.6 %).