科研之友
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摘要
This review introduces microdevices for the formation of a planar bilayer lipid membrane (BLM) array that have been developed for ion channel analyses. Cell membranes contain various types of membrane proteins that play important physiological roles, such as signal transductions, neurotransmissions, and essential nutrients/wastes transports. Accordingly, the membrane proteins are considered to be one of the most important targets for pharmaceutical drugs. Here, we focused especially on launching screening platforms for ion channel characterizations that would contribute not only to fundamental studies, but to the speed-up and cost-cutting of drug discovery. We applied the droplet contact method (DCM), one of the most simple, robust, and reproducible formation methods of a planar BLM, to replicate a biological membrane in vitro. DCM makes use of the monolayer self-assembly of amphiphilic lipid molecules at the interface between an organic solvent and an aqueous solution; BLM is formed at the interface of two contacting aqueous droplets. This principle was implemented by a microdevice that simply consists of two microwells contacting each other, taking functions of aligning the aqueous droplets and stabilizing the contact interface. Since DCM provides a sufficiently high electrical resistance of BLM, a pair of mounted Ag/AgCl electrodes enabled electrical current recordings of a single ion channel with a picoampere level. We further integrated a solution-exchanging microfluidic unit, and expanded the device to an array design for the automation and mass production of BLM, and preliminarily demonstrated ligand screenings on a nanopore membrane protein. Finally, a series of feasibility tests were performed using several types of ion channels, which proved that our platform enhances the data-throughput for the characterizations of individual ion channels, and would help to reveal the gating mechanisms of the ion channels.
- 出版日期2015-6
