Attolitre-sized lipid bilayer chamber array for rapid detection of single transporters

We present an attolitre-sized arrayed lipid bilayer chamber system (aL-ALBiC) for rapid and massively parallel single-molecule assay of membrane transporter activity. Because of the small reaction volume (200 aL), the aL-ALBiC performed fast detection of single transporter activity, thereby enhancing the sensitivity, throughput, and accuracy of the analysis. Thus, aL-ALBiC broadens the opportunities for single-molecule analysis of various membrane transporters and can be used in pharmaceutical applications such as drug screening.

[1]  P. Tinnefeld,et al.  A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research , 2014, Molecules.

[2]  Shoji Takeuchi,et al.  Microfluidic lipid membrane formation on microchamber arrays. , 2011, Lab on a chip.

[3]  R. Tampé,et al.  Multiplexed parallel single transport recordings on nanopore arrays. , 2010, Nano letters.

[4]  A. Rzhetsky,et al.  The human ATP-binding cassette (ABC) transporter superfamily. , 2001, Genome research.

[5]  M. Saier.  A Functional-Phylogenetic Classification System for Transmembrane Solute Transporters , 2000, Microbiology and Molecular Biology Reviews.

[6]  M. Niemi,et al.  Membrane transporters in drug development , 2010, Nature Reviews Drug Discovery.

[7]  Hiroyuki Noji,et al.  A single-molecule enzymatic assay in a directly accessible femtoliter droplet array. , 2010, Lab on a chip.

[8]  Hiroaki Suzuki,et al.  Ninety-six-well planar lipid bilayer chip for ion channel recording Fabricated by hybrid stereolithography , 2009, Biomedical microdevices.

[9]  H Morgan,et al.  Microfluidic array platform for simultaneous lipid bilayer membrane formation. , 2009, Biosensors & bioelectronics.

[10]  J. Gouaux,et al.  Structure of Staphylococcal α-Hemolysin, a Heptameric Transmembrane Pore , 1996, Science.

[11]  A. Barabasi,et al.  Drug—target network , 2007, Nature Biotechnology.

[12]  K. Zerf,et al.  Optical recording of signal-mediated protein transport through single nuclear pore complexes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[13]  A. Ravna,et al.  Structures and Models of Transporter Proteins , 2004, Journal of Pharmacology and Experimental Therapeutics.

[14]  S. Landfear.  Transporters for Drug Delivery and as Drug Targets in Parasitic Protozoa , 2010, Clinical pharmacology and therapeutics.

[15]  B. Le Pioufle,et al.  Activity monitoring of functional OprM using a biomimetic microfluidic device. , 2012, The Analyst.

[16]  S Bhakdi,et al.  Alpha-toxin of Staphylococcus aureus. , 1991, Microbiological reviews.

[17]  K. Torimitsu,et al.  Ca2+ ion transport through channels formed by α-hemolysin analyzed using a microwell array on a Si substrate. , 2012, Biosensors & bioelectronics.

[18]  Michele Zagnoni,et al.  Miniaturised technologies for the development of artificial lipid bilayer systems. , 2012, Lab on a chip.

[19]  R. Peri,et al.  High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology , 2008, Nature Reviews Drug Discovery.

[20]  Y. Urano,et al.  Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity , 2014, Nature Communications.

[21]  Robert Tampé,et al.  Highly Parallel Transport Recordings on a Membrane-on-Nanopore Chip at Single Molecule Resolution , 2014, Nano letters.

[22]  Shoji Takeuchi,et al.  Lipid bilayers on a picoliter microdroplet array for rapid fluorescence detection of membrane transport. , 2014, Small.

[23]  H. Noji,et al.  High-throughput formation of lipid bilayer membrane arrays with an asymmetric lipid composition , 2014, Scientific Reports.

[24]  Hiroyuki Noji,et al.  Large-scale femtoliter droplet array for digital counting of single biomolecules. , 2012, Lab on a chip.

[25]  Masayuki Fujihara,et al.  Dynamics of the spontaneous formation of a planar phospholipid bilayer: A new approach by simultaneous electrical and optical measurements , 2003 .

[26]  H. Bayley,et al.  Stochastic sensors inspired by biology , 2001, Nature.

[27]  Suhasa B. Kodandaramaiah,et al.  Automated whole-cell patch clamp electrophysiology of neurons in vivo , 2012, Nature Methods.