Droplet-based artificial membrane technology for high throughput cell-free ion ch

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DESCRIPTION (provided by applicant): Ion channels pharmaceutical discovery and safety screening is slow and expensive and as a result, there are currently no high quality, high throughput assays for ion channel screening. Librede Inc. is developing alternative cell-free technologies for ion channel measurement which have the potential for higher throughput and lower cost. In preliminary work, we have demonstrated a platform for formation and measurement of artificial cell membranes that is compatible with automated motion control hardware. Measurements of these membranes show that they are able to house ion channels with properties matching the scientific literature. The initial work with this platform utilized manual positioning and assembly of the components under constant monitoring and feedback. Although this was sufficient to demonstrate the technology, it is not sufficient to demonstrate its suitability for high throughput and automation processes. Here we propose to construct a mechanical jig which will allow assembly of the membrane components "blind", simulating automated processes. With this jig, we will cycle the membrane formation apparatus, measuring the properties of the resultant artificial membranes to determine the degree of reproducibility and therefore the suitability of this process for automation and high throughput. We have designed a plate which is compatible with industry standard 384 well fluid handling systems and contains a modification of our initial membrane formation apparatus which tolerates minor mechanical positioning errors. In the proposed work, we will use the plate and ascertain whether these modifications are sufficient or require further improvement. We will cycle the apparatus over 500 times, measuring the yield of the resulting membranes and their ability to reconstitute ion channels. The processes developed will be scalable an entire 384 well plate and the mechanical jig easily adapted for use with motion control robotics, positioning our platform for Phase II adapting it for high throughput automation. PUBLIC HEALTH RELEVANCE: Measurement of ion channel interactions with drugs is a key process in drug screening, but current technologies are slow and expensive. Our team has recently developed a method for formation of an artificial cell membrane that is compatible with robotic automation. We propose here to develop tools and processes that enable the scaling and cycling of this technology resulting in high throughput ion channel measurement.