4.5 Article

Development of an On-Chip Antibiotic Permeability Assay With Single Molecule Detection Capability

Journal

IEEE TRANSACTIONS ON NANOBIOSCIENCE
Volume 17, Issue 2, Pages 155-160

Publisher

IEEE-INST ELECTRICAL ELECTRONICS ENGINEERS INC
DOI: 10.1109/TNB.2018.2809592

Keywords

Porins; single channel electrophysiology; single molecule detection; microfluidics; nanopores

Funding

  1. Innovative Medicines Joint Undertaking from the European Unio's Seventh Framework Programme under Grant FP7 [115525]
  2. EFPIA companies

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Electrophysiology is the method of choice to characterize membrane channels. In this paper, we demonstrate a patch pipette based simple miniaturization that allows performing conductance measurements on a planar lipid bilayer in a microfluidic channel. Membrane proteins were reconstituted into Giant Unilamellar Vesicles (GUVs) by electroswelling, and GUVs with a single channel insertion were patched at the tip of pipette. We applied this approach to investigate the interactions of porins from E. coli with single antibiotics, and this will potentially provide information on the permeability rates. The results of this paper suggest that this approach can be extended to the integration of several pipettes into the microfluidic channel from different positions, allowing the multiplexed recordings and also reducing the substrate consumption below mu L volumes.

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