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Electrochemical impedance spectroscopy for black lipid membranes fused with channel protein supported on solid-state nanopore

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Abstract

Black lipid membranes (BLMs) have been used for detecting single-channel activities of pore-forming peptides and ion channels. However, the short lifetimes and poor mechanical stability of suspended bilayers limit their applications in high throughput electrophysiological experiments. In this work, we present a synthetic solid-state nanopore functionalized with BLM fused with channel protein. A nanopore with diameter of ~180 nm was electrochemically fabricated in a thin silicon membrane. Folding and painting techniques were demonstrated for production of stable suspended BLMs followed by incorporation of transmembrane protein, ENaC. Membrane formation was confirmed by employing electrochemical impedance spectroscopy (EIS) in the frequency regime of 10−2–105 Hz. Results show that electrochemically fabricated solid state nanopore support resulted in excellent membrane stability, with >1 GΩ of up to 72 and 41 h for painting and folding techniques, respectively. After fusion of ENaC channel protein, the BLM exhibits the stability of ~5 h. We anticipate that such a solid-state nanopore with diameter in the range of 150–200 nm and thickness <1 µm could be a potential platform to enhance the throughput of ion-channel characterization using BLMs.

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Acknowledgments

This work was supported by the Office of the Vice President for Research and Economic Development, University of Alabama in Huntsville (UAH), Huntsville, AL. Dr. Berdiev was supported, in whole or in part, by National Institutes of Health grant R21HL085112, the University of Alabama at Birmingham (UAB) Health Services Foundation General Endowment Fund, and Nazarbayev University Social Policy Grant.

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Authors and Affiliations

  1. Electrical and Computer Engineering Department, University of Alabama in Huntsville, Huntsville, AL, 35899, USA

    Muhammad S. Khan & John D. Williams

  2. Biotechnology Science and Engineering Program, University of Alabama in Huntsville, Huntsville, AL, 35899, USA

    Noura S. Dosoky

  3. Department of Biomedical Sciences, Nazarbayev University School of Medicine, Astana, 010000, Kazakhstan

    Bakhrom K. Berdiev

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  1. Muhammad S. Khan
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  2. Noura S. Dosoky
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Correspondence to Muhammad S. Khan.

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Khan, M.S., Dosoky, N.S., Berdiev, B.K. et al. Electrochemical impedance spectroscopy for black lipid membranes fused with channel protein supported on solid-state nanopore. Eur Biophys J 45, 843–852 (2016). https://doi.org/10.1007/s00249-016-1156-8

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  • DOI: https://doi.org/10.1007/s00249-016-1156-8

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