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Storable droplet interface lipid bilayers for cell-free ion channel studies

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Abstract

An artificially created lipid bilayer is an important platform in studying ion channels and engineered biosensor applications. However, a lipid bilayer created using conventional techniques is fragile and short-lived, and the measurement of ion channels requires expertise and laborious procedures, precluding practical applications. Here, we demonstrate a storable droplet lipid bilayer precursor frozen with ion channels, resulting in a droplet interface bilayer upon thawing. A small vial with an aqueous droplet in organic solution was flash frozen in −80 °C methanol immediately after an aqueous droplet was introduced into the organic solution and gravity draws the droplet down to the interface upon thawing. A lipid bilayer created along the interface using this method had giga-ohm resistance and typical specific capacitance values. The noise level of this system is favorably comparable to the conventional system. The subsequent incorporation of ion channels, alpha-hemolysin and gramicidin A, showed typical conductance values consistent with those in previous literatures. This novel system to create a lipid bilayer as a whole can be automated from its manufacture to use and indefinitely stored when frozen. As a result, ion channel measurements can be carried out in any place, increasing the accessibility of ion channel studies as well as a number of applications, such as biosensors, ion channel drug screening, and biophysical studies.

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Abbreviations

HTS:

High-throughput screening

hERG:

Human ether-á-go-go related gene

DPhPC:

1,2-Diphytanoyl-sn-glycero-3-phosphocholine

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Acknowledgments

This research was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (#2009-0068624, #2009-0073070) and Inha University Research Grant.

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Authors and Affiliations

  1. Department of Biological Engineering, Inha University, Incheon, Korea

    Sung-Ho Jung, Sangbaek Choi & Tae-Joon Jeon

  2. Institute of Life Science and Resources and Department of Food Science and Biotechnology, Kyung Hee University, Yongin, Korea

    Young-Rok Kim

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  1. Sung-Ho Jung
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  2. Sangbaek Choi
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  4. Tae-Joon Jeon
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Correspondence to Tae-Joon Jeon.

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Jung, SH., Choi, S., Kim, YR. et al. Storable droplet interface lipid bilayers for cell-free ion channel studies. Bioprocess Biosyst Eng 35, 241–246 (2012). https://doi.org/10.1007/s00449-011-0602-3

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  • DOI: https://doi.org/10.1007/s00449-011-0602-3

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