Abstract
Essentially all biochemistry and most molecular biology experiments require recombinant proteins. However, large, hydrophobic proteins typically aggregate into insoluble and misfolded species, and are directed into inclusion bodies. Current techniques to fold proteins recovered from inclusion bodies rely on denaturation followed by dialysis or rapid dilution. Such approaches can be time consuming, wasteful, and inefficient. Here, we describe rapid protein folding using a vortex fluidic device (VFD). This process uses mechanical energy introduced into thin films to rapidly and efficiently fold proteins. With the VFD in continuous flow mode, large volumes of protein solution can be processed per day with 100-fold reductions in both folding times and buffer volumes.
*These authors contributed equally to this work.
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Acknowledgments
J.B. thanks the Taihi Hong Memorial award for support. G.W. gratefully acknowledges support from the National Institute of General Medical Sciences of the NIH (1RO1-GM100700-01). C.R. acknowledges the Australian Research Council and the Government of South Australia for their financial support of this project.
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Britton, J., Smith, J.N., Raston, C.L., Weiss, G.A. (2017). Protein Folding Using a Vortex Fluidic Device. In: Burgess-Brown, N. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 1586. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6887-9_13
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DOI: https://doi.org/10.1007/978-1-4939-6887-9_13
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