Automated high-throughput experimentation for a fast and reliable bioprocess development

  • Automatisiertes Experimentieren im Hochdurchsatz für eine schnelle und zuverlässige Bioprozessentwicklung

Kunze, Martin; Büchs, Jochen (Thesis advisor); Jaeger, Karl-Erich (Thesis advisor)

Aachen (2019, 2020)
Dissertation / PhD Thesis

Dissertation, Rheinisch-Westfälische Technische Hochschule, 2019

Abstract

The demand for feasible bioprocesses is continuously increasing in order to realize new production pathways, convert yet unconsidered raw materials, and allow new or advanced product portfolios. One bioprocess of major interest over the last years is the production of fuels or chemicals from biomass. One key process within this production chain is the degradation of recalcitrant cellulose to better processible soluble sugars, and cellulases are key players in the hydrolyzation of cellulose. To identify best working cellulase candidates and their production hosts a reliable screening procedure is necessary. Therefore, a robotic platform is presented, the so called “Cellulolytic RoboLector”. Automated upstream, downstream, and analytical process steps in high-throughput are described and evaluated using different microbial and enzymatic candidates as model systems. One key technique within the RoboLector system, the BioLector device, allows for high-throughput online monitoring of process relevant parameters via optical methods. During the extensive work with the BioLector, some yet undiscovered pitfalls were identified which might mislead the user of such techniques and cause incorrect results. Solutions are presented to overcome these obstacles either by mathematical correction or experimental modifications. Finally, a unique system for rapid determination of temperature dependent reaction kinetics for microbial and enzymatic reactions is presented. The combination of an optical on-line monitoring device with a customized temperature control unit for 96 well microtiter plates allows high-throughput temperature profiling. Taking altogether, this work gives an extensive overview how novel automated high-throughput protocols for bioprocess development and characterization are designed, realized, and evaluated.

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