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Optical Waveguide Light-Mode Spectroscopy for Ion Channel Profiling

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Label-Free Biosensor Methods in Drug Discovery

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

Abstract

Ion channel-based biosensors using label-free optical waveguide light-mode spectroscopy (OWLS) technique provide a sensitive measurement method of trans-channel ion transport, and allow further development in utilization of ion channels as models for pharmacological purposes (drug design targeting ion channels or diagnostic applications in clinical trials). This chapter describes a sensor setup for supported cell-derived membrane fragments deposited onto a hydrophilic polytetrafluoroethylene membrane with further separation from the OWLS sensor surface by a thin polyethylene terephthalate membrane. This approach provides spatial separation between the lipid layer and the sensor surface, and also allows space for possible extramembranous domains of the inbuilt membrane channel proteins. Influx of Cl− ions through GABAA channels in the presence or absence of GABA and channel blocking agent bicuculline is measured by changes of the optical characteristics in the evanescent field at near proximity of the OWLS sensor surface.

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Acknowledgments

The author expresses her sincere appreciation to her coworkers in the study that resulted in the original publication related to this protocol report. Particular thanks are due to Emilia Madarász (Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary), István Szendrő and Katalin Erdélyi (Microvacuum Ltd., Budapest, Hungary), Pál Gróf and Nóra Kaszás (Semmelweis University, Budapest, Hungary), as well as Ferenc A. Anthony, Balázs Mihalik, and Ágnes Pataki (EGIS Pharmaceutical Co., Budapest, Hungary) for their contribution, technical support, and helpful discussions in the OWLS technique, liposome preparation, and HEK293 cell line, expressing GABAA (α5, β2, γ2) receptors, respectively. The material support by Oxyphen GmbH (Zürich, Switzerland) by providing samples of RoTrack membranes is also acknowledged.

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Authors and Affiliations

  1. Institute of Technical Physics and Materials Science, Centre for Energy Research, Hungarian Academy of Sciences, Konkoly-Thege Miklós út 29-33, Budapest, 1121, Hungary

    Inna Székács

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  1. Inna Székács
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Correspondence to Inna Székács .

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Editors and Affiliations

  1. Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, New York, USA

    Ye Fang

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Székács, I. (2015). Optical Waveguide Light-Mode Spectroscopy for Ion Channel Profiling. In: Fang, Y. (eds) Label-Free Biosensor Methods in Drug Discovery. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2617-6_8

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  • DOI: https://doi.org/10.1007/978-1-4939-2617-6_8

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2616-9

  • Online ISBN: 978-1-4939-2617-6

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