Development and characterisation of a new fluorescence sensor for online monitoring of bioprocesses
2018
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 µmol L −1 for NADH
and 0.0025–7.5 µmol L −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 %).
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