Lipid Bilayers at Gel/Gel Interface for Ion Channel Recordings

Hirano, Minako; Kobayashi, Toshihide; Ide, Toru

doi:10.1380/ejssnt.2008.130

Cited by 5 publications

(8 citation statements)

References 14 publications

(9 reference statements)

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“…It appears that organic solvent or excess lipid was present in the thick lipid layer and that the two lipid monolayers contacted each other to form the bilayer. Furthermore, it has been observed that bilayer formation proceeds until the membrane forms regardless of the interface. − …”

Section: Resultsmentioning

confidence: 99%

Current Recordings of Ion Channel Proteins Immobilized on Resin Beads

et al. 2009

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Current ion channel current measurement techniques are cumbersome, as they require many steps and much time. This is especially true when reconstituting channels into liposomes and incorporating them into lipid bilayers. Here, we report a novel method that measures ion channel current more efficiently than current methods. We applied our method to KcsA and MthK channels by binding them to cobalt affinity gel beads with histidine tags and then forming a lipid bilayer membrane on the bead. This allowed channels to incorporate into the bilayer and channel currents to be measured quickly and easily. The efficiency was such that currents could be recorded with extremely low amounts of protein. In addition, the channel direction could be determined by the histidine tag. This method has the potential to be applied to various channel proteins and channel research in general.

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Section: Resultsmentioning

confidence: 99%

Current Recordings of Ion Channel Proteins Immobilized on Resin Beads

et al. 2009

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“…There have been a number of recent developments in artificial bilayer platforms with improved robustness and longevity, microfluidic integration, and parallelization, which indicate the potential of this technology to widen its range of applicability. [5][6][7][8][9][10] In particular, artificial bilayers formed through mechanical contact of lipid monolayers at aqueous/oil interfaces or droplets [11][12][13][14] have enabled implementation in microfluidic devices, 15,16 automation, [17][18][19] and arrays. [20][21][22] These developments in artificial bilayer platforms have the potential to make practical their use in pharmaceutical ion channel screening, alongside automated and parallelized patch clamp platforms.…”

Section: Introductionmentioning

confidence: 99%

Ion channel drug potency assay with an artificial bilayer chip

2012

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The potency of pharmaceutical compounds acting on ion channels can be determined through measurements of ion channel conductance as a function of compound concentration. We have developed an artificial lipid bilayer chip for simple, fast, and high-yield measurement of ion channel conductance with simultaneous solution perfusion. Here we show the application of this chip to the measurement of the mammalian cold and menthol receptor TRPM8. Ensemble measurements of TRPM8 as a function of concentration of menthol and 2-aminoethoxydiphenyl borate (2-APB) enabled efficient determination of menthol's EC(50) (111.8 μM ± 2.4 μM) and 2-APB's IC(50) (4.9 μM ± 0.2 μM) in agreement with published values. This validation, coupled with the compatibility of this platform with automation and parallelization, indicates significant potential for large-scale pharmaceutical ion channel screening.

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“…When measuring channel currents of ion channels reconstituted into the solid-supported lipid bilayers, ionic currents passing through membrane proteins and peptides are typically lower than their original value since the lipid bilayer is in direct contact with the substrate blocking ion flow . To overcome this issue, the use of porous membranes − or hydrogel layers − as the substrate and the addition of a hydrophilic layer such as hydrogel or polymer layer have previously been reported . The porous-membrane-supported lipid bilayer is achieved via the use of small pores; it typically requires cumbersome fabrication skills.…”

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confidence: 99%

Recessed Ag/AgCl Microelectrode-Supported Lipid Bilayer for Nanopore Sensing

2020

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Biological nanopores reconstituted into supported lipid bilayer membranes are widely used as a platform for stochastic nanopore sensing with the ability to detect single molecules including, for example, single-stranded DNA (ssDNA) and miRNA. A main thrust in this area of research has been to improve overall bilayer stability and ease of measurements. These improvements are achieved through a variety of clever strategies including droplet-based techniques; however, they typically require specific microfabrication techniques to prepare devices or special manipulation techniques for microdroplets. Here, we describe a new method to prepare lipid bilayers using a recessed-in-glass Ag/AgCl microelectrode as a support structure. The lipid bilayer is formed at the tip of the microelectrode by immersing the microelectrode into a layered bath solution consisting of an oil/lipid mixture and an aqueous electrolyte solution. In this paper, we demonstrate this stable, supported lipid bilayer structure for channel current measurements of pore-forming toxins and single-molecule detection of ssDNA. This Ag/AgCl-supported lipid bilayer can potentially be widely adopted as a lipid membrane platform for nanopore sensing because of its simple and easy procedure needed to prepare lipid bilayers.

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Lipid Bilayers at Gel/Gel Interface for Ion Channel Recordings

Cited by 5 publications

References 14 publications

Current Recordings of Ion Channel Proteins Immobilized on Resin Beads

Current Recordings of Ion Channel Proteins Immobilized on Resin Beads

Ion channel drug potency assay with an artificial bilayer chip

Recessed Ag/AgCl Microelectrode-Supported Lipid Bilayer for Nanopore Sensing

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