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Membrane-on-a-Chip: Microstructured Silicon/Silicon-Dioxide Chips for High-Throughput Screening of Membrane Transport and Viral Membrane Fusion

Kusters, I., van Oijen, A. M. & Driessen, A. J. M., Apr-2014, In : Acs Nano. 8, 4, p. 3380-3392 13 p.

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  • Membrane-on-a-Chip Microstructured SiliconSilicon-Dioxide Chips for High-Throughput Screening of Membrane Transport and Viral Membrane Fusion

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DOI

Screening of transport processes across biological membranes is hindered by the challenge to establish fragile supported lipid bilayers and the difficulty to determine at which side of the membrane reactants reside. Here, we present a method for the generation of suspended lipid bilayers with physiological relevant lipid compositions on microstructured Si/SiO2 chips that allow for high-throughput screening of both membrane transport and viral membrane fusion. Simultaneous observation of hundreds of single-membrane channels yields statistical information revealing population heterogeneities of the pore assembly and conductance of the bacterial toxin a-hemolysin (alpha HL). The influence of lipid composition and ionic strength on aHL pore formation was investigated at the single-channel level, resolving features of the pore-assembly pathway. l Pore formation is inhibited by a specific antibody, demonstrating the applicability of the platform for drug screening of bacterial toxins and cell-penetrating agents. Furthermore, fusion of H3N2 influenza viruses with suspended lipid bilayers can be observed directly using a specialized chip architecture. The presented micropore arrays are compatible with fluorescence readout from below using an air objective, thus allowing high-throughput screening of membrane transport in multiwell formats in analogy to plate readers.

Original languageEnglish
Pages (from-to)3380-3392
Number of pages13
JournalAcs Nano
Volume8
Issue number4
Publication statusPublished - Apr-2014

    Keywords

  • micropore array, suspended lipid bilayers, membrane transport, virus fusion, alpha-hemolysin, STAPHYLOCOCCAL ALPHA-HEMOLYSIN, GIANT UNILAMELLAR VESICLES, HEPTAMERIC TRANSMEMBRANE PORE, PHOSPHOLIPID-BILAYERS, FORMING PROTEIN, HIGH-RESOLUTION, TOXIN, ELECTROFORMATION, RECONSTITUTION, STOICHIOMETRY

ID: 13789026