Detecting proteins complex formation using steady-state diffusion in a nanochannel |
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Authors: | Nicolas F Y Durand Elli Saveriades Philippe Renaud |
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Institution: | (1) Microsystems Laboratory, STI-LMIS, EPFL, 1015 Lausanne, Switzerland |
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Abstract: | In this work, we present theoretical and experimental studies of nanofluidic channels as a potential biosensor for measuring
rapid protein complex formation. Using the specific properties offered by nanofluidics, such as the decrease of effective
diffusion of biomolecules in confined spaces, we are able to monitor the binding affinity of two proteins. We propose a theoretical
model describing the concentration profile of proteins in a nanoslit and show that a complex composed by two bound biomolecules
induces a wider diffusion profile than a single protein when driven through a nanochannel. To validate this model experimentally,
we measured the increase of the fluorescent diffusion profile when specific biotinylated dextran was added to fluorescent
streptavidin. We report here a direct and relatively simple technique to measure the affinity between proteins.
Figure We present theoretical and experimental studies of nanofluidic channels as potential biosensors for rapidly measuring protein
complex formation. Our system is based on steady-state diffusion effects which are observed inside a nanoslit. |
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Keywords: | Bioassay Nanofluidics Nanochannel Fluorescence Proteomics Microfabrication |
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