Delivery of synthetic antibodies to probe cell dynamics in live cells

Type of Award: Spark
Date Awarded: September 2009
Award End Date: August 2011
Amount Awarded: $ 400,000.00
PI(s): Anthony Kossiakoff, PhD, UChicago; Vladimir Gelfand, PhD, NU; Charles Clevenger, PhD, NU;

Abstract: Goals: We propose to develop a technology platform for the next generation of affinity reagents and imaging probes that will substantially extend capabilities for observing dynamic events in live cells. Transitions between distinct protein conformations or the formation of multi-protein complexes are fundamental to cellular structures, signaling and regulation. However, current antibody and fluorescent-protein tags are generally insensitive to these subtle, but functionally critical changes in target molecules. To overcome this deficiency we have developed a new technology platform that both generates highly specific affinity-binders that have the capability to identify specific protein complexes and transient conformations and to deliver these probes to living cells with experimental ease and minimal perturbation so they can interrogate cellular dynamics in their native cell environments. As a challenging test-bed to develop and test our new technology, we will generate unique affinity reagents and imaging probes to study core aspects of signaling and motor function essential in cytoskeletal dynamics that cannot be investigated using current methods. This pilot project is only the beginning for what we believe is the full potential for our technology platform. Our long term objective is to assemble a high-throughput pipeline to produce high-affinity synthetic affinity reagents to high-impact protein and RNA targets for use in imaging, localization and inhibition studies in their native cellular environments. We believe it is well within the capability of the technology to automate most of the steps of the pipeline with the aim to create a powerful, compact and affordable core facility-like platform to provide all Chicago area researchers with designer antibodies that can be tailored to particular systems and needs.