BioVT-2010-18 [BibTeX]
Matthias Funke, Andreas Buchenauer, Wilfried Mokwa, Stefanie Kluge, Lea Hein, Carsten Müller, Frank Kensy, Jochen Büchs:
Bioprocess control in Microscale: Scalable fermentations in disposable and user-friendly microfluidic systems
Microbial Cell Factories, 2010, 9:86
Abstract:
Background: The efficiency of biotechnological production processes depends on selecting the best performing
microbial strain and the optimal cultivation conditions. Thus, many experiments have to be conducted, which
conflicts with the demand to speed up drug development processes. Consequently, there is a great need for highthroughput
devices that allow rapid and reliable bioprocess development. This need is addressed, for example, by
the fiber-optic online-monitoring system BioLector which utilizes the wells of shaken microtiter plates (MTPs) as
small-scale fermenters. To further improve the application of MTPs as microbioreactors, in this paper, the BioLector
technology is combined with microfluidic bioprocess control in MTPs. To realize a user-friendly system for routine
laboratory work, disposable microfluidic MTPs are utilized which are actuated by a user-friendly pneumatic
hardware.
Results: This novel microfermentation system was tested in pH-controlled batch as well as in fed-batch
fermentations of Escherichia coli. The pH-value in the culture broth could be kept in a narrow dead band of 0.03
around the pH-setpoint, by pneumatically dosing ammonia solution and phosphoric acid to each culture well.
Furthermore, fed-batch cultivations with linear and exponential feeding of 500 g/L glucose solution were
conducted. Finally, the scale-up potential of the microscale fermentations was evaluated by comparing the
obtained results to that of fully controlled fermentations in a 2 L laboratory-scale fermenter (working volume of 1 L).
The scale-up was realized by keeping the volumetric mass transfer coefficient kLa constant at a value of 460 1/h. The
same growth behavior of the E. coli cultures could be observed on both scales.
Conclusion: In microfluidic MTPs, pH-controlled batch as well as fed-batch fermentations were successfully
performed. The liquid dosing as well as the biomass growth kinetics of the process-controlled fermentations
agreed well both in the microscale and laboratory scale. In conclusion, a user-friendly and disposable microfluidic
system could be established which allows scaleable, fully controlled and fully monitored fermentations in working
volumes below 1 milliliter.
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