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Design and Application of Software Sensors in Batch and Fed-batch Cultivations during Recombinant Protein Expression in Escherichia coli
Linköping University, The Department of Physics, Chemistry and Biology. (Teknisk Biologi)
2008 (English)Independent thesis Advanced level (degree of Master), 20 points / 30 hpStudent thesis
Abstract [en]

Software sensors are a potent tool to improve biotechnological real time process monitoring and control. In the current project, algorithms for six partly novel, software sensors were established and tested in a microbial reactor system. Eight batch and two fed-batch runs were carried out with a recombinant Escherichia coli to investigate the suitability of the different software sensor models in diverse cultivation stages. Special respect was given to effects on the sensors after recombinant protein expression was initiated by addition of an inducer molecule. It was an objective to figure out influences of excessive recombinant protein expression on the software sensor signals.

Two of the developed algorithms calculated the biomass on-line and estimated furthermore, the specific growth rate by integration of the biomass changes with the time. The principle of the first was the application of a near infrared probe to obtain on-line readings of the optical density. The other algorithm was founded on the titration of ammonia as only available nitrogen source. The other two sensors analyzed for the specific consumption of glucose and the specific production of acetate and are predicted on an in-line HPLC system.

The results showed that all software sensors worked as expected and are rather powerful to estimate important state parameters in real time. In some stages, restrictions may occur due to different limitation affects in the models or the physiology of the culture. However, the results were very convincing and suggested the development of further and more advanced software sensor models in the future.

Place, publisher, year, edition, pages
2008. , 62 p.
Software sensor, Escherichia coli, Green fluorescence protein, Process analytical technology (PAT), Biomass, Metabolic stress
National Category
Industrial Biotechnology
URN: urn:nbn:se:liu:diva-12530ISRN: LiTH-IFM-A-EX-08/2013-SEOAI: diva2:406
Available from: 2008-10-14 Created: 2008-09-11 Last updated: 2008-10-14Bibliographically approved

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