7 December 2013 Integrated microdroplet-based system for enzyme synthesis and sampling
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Abstract
Microdroplet-based microfluidic devices are emerging as powerful tools for a wide range of biochemical screenings and analyses. Monodispersed aqueous microdroplets from picoliters to nanoliters in volume are generated inside microfluidic channels within an immiscible oil phase. This results in the formation of emulsions which can contain various reagents for chemical reactions and can be considered as discrete bioreactors. In this paper an integrated microfluidic platform for the synthesis, screening and sorting of libraries of an organophosphate degrading enzyme is presented. The variants of the selected enzyme are synthesized from a DNA source using in-vitro transcription and translation method. The synthesis occurs inside water-in-oil emulsion droplets, acting as bioreactors. Through a fluorescence based detection system, only the most efficient enzymes are selected. All the necessary steps from the enzyme synthesis to selection of the best genes (producing the highest enzyme activity) are thus integrated inside a single and unique device. In the second part of the paper, an innovative design of the microfluidic platform is presented, integrating an electronic prototyping board for ensuring the communication between the various components of the platform (camera, syringe pumps and high voltage power supply), resulting in a future handheld, user-friendly, fully automated device for enzyme synthesis, screening and selection. An overview on the capabilities as well as future perspectives of this new microfluidic platform is provided.
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Florian Lapierre, Florian Lapierre, Michel Best, Michel Best, Robert Stewart, Robert Stewart, John Oakeshott, John Oakeshott, Thomas Peat, Thomas Peat, Yonggang Zhu, Yonggang Zhu, } "Integrated microdroplet-based system for enzyme synthesis and sampling", Proc. SPIE 8923, Micro/Nano Materials, Devices, and Systems, 89233C (7 December 2013); doi: 10.1117/12.2033569; https://doi.org/10.1117/12.2033569
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