Refolding of β-galactosidase: microfluidic device for reagent metering and mixing and quantification of refolding yield |
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Authors: | Sowmya Kondapalli Brian J Kirby |
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Affiliation: | (1) Sibley School of Mechanical and Aerospace Engineering, Cornell University, Ithaca, NY 14853, USA |
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Abstract: | We have developed a device that uses microfluidic valves and pumps to meter reagents for subsequent mixing with application
to refolding of the protein β-galactosidase. The microfluidic approach offers the potential advantages of automation, cost-effectiveness,
compatibility with optical detection, and reduction in sample volumes as opposed to conventional techniques of hand-pipetting
or using robotic systems. The device is a multi-layered poly(dimethylsiloxane) on glass device with automated controls for
reagent aliquoting and mixing. Refolding experiments have been performed off-chip using existing protocols on the protein
β-galactosidase and the refolding yield has been quantified on-chip using fluorescein di-β-d-galactopyranoside, a caged-fluorescent molecule. This work provides the potential to reduce the cost of drug discovery and
realization of protein pharmaceuticals. |
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Keywords: | Protein refolding β -Galactosidase Combinatorial Microfluidic |
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