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Monitoring and Quantifying the Passive Transport of Molecules Through Patch–Clamp Suspended Real and Model Cell Membranes
Authors:Dr Pierluca Messina  Dr Frédéric Lemaître  Prof François Huet  Dr Kieu An Ngo  Dr Vincent Vivier  Prof Eric Labbé  Dr Olivier Buriez  Prof Christian Amatore
Institution:1. Ecole Normale Supérieure, Département de Chimie, UMR CNRS‐ENS‐UPMC 8640 “PASTEUR”, 24 rue Lhomond, 75231 Paris cedex 05 (France);2. LISE, CNRS UPR 15, Université Pierre et Marie Curie, CP 133, 4 Place Jussieu, 75252 Paris Cedex 05 (France)
Abstract:Transport of active molecules across biological membranes is a central issue for the success of many pharmaceutical strategies. Herein, we combine the patch–clamp principle with amperometric detection for monitoring fluxes of redox‐tagged molecular species across a suspended membrane patched from a macrophage. Solvent‐ and protein‐free lipid bilayers (DPhPC, DOPC, DOPG) patched from single‐wall GUV have been thoroughly investigated and the corresponding fluxes measurements quantified. The quality of the patches and their proper sealing were successfully characterized by electrochemical impedance spectroscopy. This procedure appears versatile and perfectly adequate to allow the investigation of transport and quantification of the transport properties through direct measurement of the coefficients of partition and diffusion of the compound in the membrane, thus offering insight on such important biological and pharmacological issues.
Keywords:amperometry  membranes  patch–  clamp  transport  ultra‐microelectrode
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