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Polymer-based microfluidic device for measuring membrane protein activities

Hutter, I. ; Müller, E. ; Kristiansen, P. ; Kresak, S. ; Tiefenauer, L.

In: Microfluidics and Nanofluidics, 2013, vol. 14, no. 3-4, p. 421-429

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    Titre
    • Polymer-based microfluidic device for measuring membrane protein activities
    Auteur
    • Hutter, I.. Paul Scherrer Institute (PSI), Biomolecular Research (BMR), 5232, Villigen, Switzerland
    • Müller, E.. Institute of Polymer Nanotechnology, University of Applied Sciences and Arts FHNW, 5210, Windisch, Switzerland
    • Kristiansen, P.. Institute of Polymer Nanotechnology, University of Applied Sciences and Arts FHNW, 5210, Windisch, Switzerland
    • Kresak, S.. Max Planck Institute of Biophysics (MPIBP), 60438, Frankfurt a.M., Germany
    • Tiefenauer, L.. Paul Scherrer Institute (PSI), Biomolecular Research (BMR), 5232, Villigen, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Microfluidics and Nanofluidics, 2013, vol. 14, no. 3-4, p. 421-429. Springer-Verlag
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:313889
    Summary
    Functional assays of membrane proteins are becoming increasingly important, both in research and drug discovery applications. The majority of current assays use the patch-clamp technology to measure the activity of ion channels which are over-expressed in cells. In future, in vitro assay systems will be available, which use reconstituted membrane proteins in free-standing lipid bilayers suspended in nano- or micrometer-sized pores. Such functional assays require (1) expression, purification and reconstitution of the membrane protein of interest, (2) a reliable method for lipid bilayer formation and membrane protein integration, and (3) a sensitive detection system. For practical applications, especially for automation, the reliable and controllable transport of fluids is essential. In order to achieve a stable free-standing lipid bilayer, a pore diameter in the micro- to nanometer range is essential. Novel microfluidic devices were developed by bonding a thick (300μm) polyether ether ketone foil, bearing a channel structure, to a thin (12μm) foil with a micropore of about 10μm diameter and then utilized for the formation of stable, free-standing lipid bilayers within the pore. A bacterial voltage-gated potassium channel is integrated therein by fusion and the ion channel activity detected by voltage clamp