Hybrid biological/artificial nanopore
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1 Hybrid biological/artificial nanopore Sébastien Balme, Simon Cabello Aguilar, Mathilde Lepoitevin, Mikhael Bechelany Adib Abou Chaaya, Jean Marc Janot, Emmanuel Balanzat, Philippe Déjardin Emir user days, Orsay, October 2013
2 Micrométrique Analogy between biological and Biological Nanopore artificial membrane Membrane protein Delicate 10 nm High selectivity Solid state Nanopore Nanopore Polymère par irradiation Nitrure de silice Graphène Design (shape size number) Robusness Poor selectivity Cylindrique Conique Biconique Bicylindrique
3 Combine the advantages of both Solid state / Biological Robustness Number, Size and shape control Selectivity, Atomically precise structure Hybrid nanopore based on channel protein confinement Dekker s groups Hall et al. Nature Nanotechnology 5, (2010) S. Balme et al., Nano Letters, 11, 712 (2011)
4 Hybrid nanopore based on Gramicidin A confined inside commercial nanopore Biological Membrane Artificial nanopore Selectivity on monovalent cation H + > K + > Na + Head-Head Dimers No selectivity on monovalent cation K + > Na + = Ca 2+ Lost Head-Head Dimers Blocking Proton S. Balme et al., Nanoscale, 5, , (2013) S. Balme et al., Nano Letters, 11, 712 (2011) D. Bonhenry et al. Soft Matter, 7, (2011)
5 Concept for hybrid nanopore building Design Nanopore (shape, size, number) a diameter close to the hydrophobic barrel of the membrane protein surface functions which permit to mimic phospholipid chain / protein barrel interactions a geometry which allows protein relaxation during transport process Biological channel activity Proteins confinement Localisation, structure Recovery of channel properties? Biological activity (transport, selectivity) Fonctionnal Hybrid biological/artificial nanopore
6 COO- COO- COO- COO- COO- COO- COO- Single nanopore design Kr 35+ Cimap - GANIL Caen with E. Balanzat Experiments P923-M PET film (µm) «Etching» NaOH 3M 50 C d > 20 nm Carboxylate function Nanopore chargé 1 nm < d < 10 nm C H 3 CH 3 Si CH 3 C H 3 CH 3 Si CH 3 10 mn SH SH ZnO/Al 2 O 3 -TMS 30 mn Atomic Layer Deposition (ALD) ** Hydrophobic Nanopore
7 Ionics transport mechanism through confined ga With ga Λ out = With ga 1.2 With ga A nm Without ga 0.2 Without ga 0.2 Without ga Without ga (A) [NaCl] (M) (B) [NaCl] (M) [NaCl] (M) [NaCl] (M) (B) (C) With ga With ga 2 nm Without ga 0.0 Without ga (C) [NaCl] (M) [NaCl] (M) G T = g Max Na + Na+ K M Na + + Na + + g Max Cl If Nanopore diameter 6 Cl K M Cl + Cl + λ out[nacl] mn ionic transport is only inside ga A. Abou Chaaya et al. J. Phys Chem C, 117 (29) (2013)
8 Nanopore DNA biosensor α-hemolysin DNA translocation Discrimination nucleotid Fast translocation 2µs.base -1 Barrel ~4 nm Hydrophylic part 10 nm Applications : DNA sequencing, diagnostic, food analysis Deamer et al. Trends in biotechnology, 18, , 2000
9 (a) Recovery of biological properties ZnO / Al2O3 HMDS (a) (b) 300 Poly(A) Poly(A) Poly(C) 300 Poly(C) Events (counts) 200 Poly(U) Poly(U) Poly(I) 300 I/I0 (b) (b) Cabello-Aguilar et al. Nanoscale, 2013,5, Poly(I)
10 Conclusion Ionic channel (a) Recovery of biological properties Nucleotide discrimination Poly(A) Poly(C) Poly(U) (b) Recovery of ionic transport mechanism Selectivity is lost Poly(I) Hemolysin Feasibility to combine the advantages of both solid state and biology
11 Futur for hybrid nanopores Water desalination DNA Sequencing Aquaporins Dialysis Diagnostic Hemolysin MSPA PHI 29 Nanopores Ionic channels «Phenotyping» Filtration Receptor channels Diagnostic tools Bio-sensor Drug testing
12 Aknowledgments IEM, Montpellier S. Cabello-Aguillar M. Lepoitevin A. Abou-Chaaya ICSM, Marcoule Dr. J Cabedouzou CIMAP-GANIL, Caen E. Balanzat Financial supports and Facilities Dr. P. Déjardin Dr. J.-M. Janot Dr. M. Bechelany Proposal Experiments P923-M Cimap-GANIL Grant SIMI 8 TRANSION Post-doc position will be opened in 2014 Topic «Hybrid ionic channel / artificial Nanopore» Thank you for your attention
13 Control nanopore size and shape using Atomic Layer deposition Conformal coating One Cycle of ALD: Step 1 :Pulse precursor 1 Step 2 : Exposure + Purge Step 3 : Pulse precursor 2 Step 4 : Exposure + Purge Theoretically ->1 monolayer
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