Electrophysiology of Ion channels in miniaturised systems

Cell membranes form a natural protective boundary around cells and their organelles. Ion channels housed within these membranes comprise 1.5% of the human genome and carry out essential cell signalling roles. As such, ion channels are important pharmacological targets and a better understanding of their function would aid drug discovery as well as drug toxicity testing. Although there are methods of studying ion channels, including patch clamping and artificial lipid bilayer system, a number of difficulties limit their experimental efficiency and practicality. In the case of artificial bilayer architectures, reconstitution of membrane proteins into bilayers is challenging. Here we demonstrate the design and development of a scalable droplet interface bilayer system for single ion channel electrophysiology. A combination of this platform and the ion channel reconstitution method addresses the protein integration problem by improving the probability of channel incorporation to 29%. Single channel recordings of gramicidin, alamethicin and alpha hemolysin were acquired for proof of concept work while eukaryotic ion channel BK electrophysiology provides evidence of the success of the project objectives. We anticipate that the suggested reconstitution method in conjunction with the platform developed in this research can be extended to study other pharmacologically relevant human ion channels.Cell membranes form a natural protective boundary around cells and their organelles. Ion channels housed within these membranes comprise 1.5% of the human genome and carry out essential cell signalling roles. As such, ion channels are important pharmacological targets and a better understanding of their function would aid drug discovery as well as drug toxicity testing. Although there are methods of studying ion channels, including patch clamping and artificial lipid bilayer system, a number of difficulties limit their experimental efficiency and practicality. In the case of artificial bilayer architectures, reconstitution of membrane proteins into bilayers is challenging. Here we demonstrate the design and development of a scalable droplet interface bilayer system for single ion channel electrophysiology. A combination of this platform and the ion channel reconstitution method addresses the protein integration problem by improving the probability of channel incorporation to 29%. Single channel recordings of gramicidin, alamethicin and alpha hemolysin were acquired for proof of concept work while eukaryotic ion channel BK electrophysiology provides evidence of the success of the project objectives. We anticipate that the suggested reconstitution method in conjunction with the platform developed in this research can be extended to study other pharmacologically relevant human ion channels