Bioprocess Control in Microscale: Scalable Fermentations in Disposable and User-Friendly Microfluidic Systems

Microbial Cell Factories

Research

Matthias Funke¹  Lea Hein¹  Jochen Büchs¹  Stefanie Kluge¹  Andreas Buchenauer²  Wilfried Mokwa²  Carsten Müller³  Frank Kensy³ 

[1] AVT-Biochemical Engineering, RWTH Aachen University, Worringerweg 1, D-52074, Aachen, Germany;Institute of Materials in Electrical Engineering 1, RWTH Aachen University, Sommerfeldstr. 24, D-52074, Aachen, Germany;m2p-labs GmbH, Forckenbeckstraße 6, D-52074, Aachen, Germany;

关键词: Fermentation;    Microfluidic Chip;    Stir Tank Reactor;    Dissolve Oxygen Tension;    Pump Chamber;   

DOI  :  10.1186/1475-2859-9-86

 received in 2010-07-12, accepted in 2010-11-13,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundThe 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 high-throughput 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.ResultsThis 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.ConclusionIn 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.

【 授权许可】

Unknown   
© Funke et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.