Drug discovery from venoms using miniaturized artificial bilayers

The endpoint of the VenomPicoScreen project is the identification of new candidate drugs acting on ion channels and receptors from venom peptides. To achieve this goal, we propose to develop a membrane protein platform for the screening of nanoliter volumes of venoms, enabling assays based on a single spider, the tiny volume of natural venom sources being the bottleneck in these investigations. This platform will bring together 4 state-of-the-art technologies: 1) DIB for 'Droplet interface bilayers' to create artificial lipid bilayers from tiny 0.3 µL droplets; 2) In-droplet expression of membrane proteins from cDNA by incorporation within droplets of in-vitro transcription/translation mix; 3) EWOD for 'Electrowetting on dielectric' to remotely manipulate multiple droplets and form lipid bilayers; 4) ICCR for 'Ion-channel coupled receptors' to transduce ligand binding to a single receptor into a detectable electrical signal. Including experts in venoms, microfluidics, electronics, electrophysiology, and protein engineering, the consortium possesses hands-on experience in each of the technologies involved. The platform will integrate on a single chip the microfluidics and electronics necessary for generating droplets, forming bilayers, and recording transbilayer currents. Tests will be directed towards proteins for which few drugs are available: initially, voltage-dependent potassium channels, and, at a later stage, selected G protein coupled receptors. Positive outcome, in addition to the advent of a new venom screening method, will be judged by the identification of venom peptides with modulatory activity on these targets.