Evaluation of software sensors for on-line estimation of culture conditions in an Escherichia coli cultivation expressing a recombinant protein.
Software sensors for monitoring and on-line estimation of critical bioprocess variables have mainly been used with standard bioreactor sensors, such as electrodes and gas analyzers, where algorithms in the software model have generated the desired state variables. In this article we propose that other on-line instruments, such as NIR probes and on-line HPLC, should be used to make more reliable and flexible software sensors. Five software sensor architectures were compared and evaluated: (1) biomass concentration from an on-line NIR probe, (2) biomass concentration from titrant addition, (3) specific growth rate from titrant addition, (4) specific growth rate from the NIR probe, and (5) specific substrate uptake rate and by-product rate from on-line HPLC and NIR probe signals. The software sensors were demonstrated on an Escherichia coli cultivation expressing a recombinant protein, green fluorescent protein (GFP), but the results could be extrapolated to other production organisms and product proteins. We conclude that well-maintained on-line instrumentation (hardware sensors) can increase the potential of software sensors. This would also strongly support the intentions with process analytical technology and quality-by-design concepts.
Warth
Benedikt
B
Division of Biotechnology/IFM, Linköping University, 581 83 Linköping, Sweden.
Rajkai
György
G
Mandenius
Carl-Fredrik
CF
D018533:Biomass; D015374:Biosensing Techniques / Q000295:instrumentation*; D018929:Cell Culture Techniques / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D009862:Online Systems / Q000295:instrumentation*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012984:Software; D013379:Substrate Specificity
D011994:Recombinant Proteins