Protein expression in microsystems
Ion channels are membrane proteins of interest to medical research, drug discovery, and biosensing applications. Expressing ion channels and inserting them into lipid bilayers for characterisation using electrophysiology is conventionally a multi-step process involving the growth and transformation of cell lines followed by cell lysis, protein purification and reconstitution. This is a labour intensive, time consuming and cumbersome process that is often limited by low yields. In vitro transcription/translation is a fast, cell-free and commercially available approach to expressing proteins. A cell-free expression mixture contains all the necessary components for expressing proteins from a supplied DNA template. One drawback of this approach is that commercial cell-free systems are expensive, which has restricted their use to a small number of specific applications. However this is not an issue for lab-on-chip technology, where sample volumes are reduced to the microliter scale.
The aim of this project is to simultaneously express and characterise ion channels on-chip inside microdroplets using in vitro transcription/translation and electrophysiology. This is achieved using a droplet dielectrophoresis device capable of forming lipid bilayers by manipulating two microdroplets into contact inside a well that contains a lipid-oil solution. This transforms the conventional multi-day, multi-step single ion-channel electrophysiology method into a quick and economical process.
- cell-free protein expression
- digital microfluidics
- ion channels
- single-channel electrophysiology
- Mark Friddin
- Prof. Hywel Morgan
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