Tailoring nanopores for efficient sensing of different biomolecules
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Tailoring nanopores for efficient sensing of different biomolecules G. Oukhaled1,2, L. Bacri2, E. Bourhis1, B. Schiedt1,2, A. Madouri1, G. Patriarche1, R. Jede3, JM. Betton5, P. Guegan2, L. Auvray2, J. Pelta2, 4, and J. Gierak1 1
LPN/CNRS, Route de Nozay, F-91460 Marcoussis, France MPI, LAMBE, Université d’Évry Val d’Essonne, Bd. François Mitterrand, F-91025 Évry, France 3 RAITH GmbH, Hauert 18, Technologiepark, D-44227 Dortmund, Germany 4 Université de Cergy-Pontoise, F-95302 Cergy-Pontoise, France 5 Unité de Biochimie Structurale, CNRS-URA 2185 Institut Pasteur, 75724 Paris, France Keywords: solid state nanopores, biosensor, nanofabrication, focused ion beam, DNA, proteins 2
ABSTRACT Highly Focused Ion Beams (FIB) are used to produce in one step large quantities of solid state nanopores drilled in thin dielectric films with high reproducibility and well controlled morphologies. We explore both the production of nanopores of various diameters and study their applicability to different biological molecules such as DNA, or folded and unfolded proteins, and then we compare their transport properties. We also report on the translocation of Fibronectin which an original experiment made possible is using the methodology described in this article. I. Introduction Many natural and technological processes involve the transport of colloidal objects or macromolecules through channels or pore of very small radius. One can cite the phenomenon of filtration [1], biopolymers synthesis [2] (DNA, RNA, and proteins), extranuclear transport of messenger RNA, the translocation and secretion of proteins [3] and the infection of a cell by certain viruses [4]. Mimicking artificially these processes at the level of single molecule is a major challenge. For the first time in 1996 the observation of the passage of a singlestranded RNA molecule through a nanometric protein pore inserted in a lipid bilayer, using a simple electrical technique has been reported [5]. This electrical detection method is the same as the one used on a more macroscopic scale in the Coulter counter [6]. It functions by measuring the electrical conductance of a pore that connects two solution-filled reservoirs. A particle entering the pore displaces conducting electrolyte, which leads to a transient change in the measured conductance. The magnitude and duration of the transient change are associated to the size and position of particle that caused it. Since then, several groups have been investigating many applications of macromolecules transport through biological nanopore, including ultrafast sequencing of DNA and RNA [7]. It soon became apparent that one way of extending this work on protein channels and getting around certain limitations in their use, such as the fragility of lipid membranes and protein channels, the lack of variability in the pore sizes, was to use artificial nanopores in thin solid state membranes. Current state of the art methods for nanopore production includes sculpting methods, direct drilling with focused electron (TEM) or i
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