Fluorogenic Molecular Proximity Detection Platform for Biology Research and Clini

DESCRIPTION (provided by applicant): This SBIR Phase I project involves the creation of a new fluorogenic detection platform compatible with point-of-care instrumentation, enabling more information-rich biomarkers to be deployed in the coming era of personalized medicine. Current methods for assessing protein activation, complex formation and localization require complicated methods and instrumentation not amenable to cost-effective point-of-care devices. The Fluorogen Activating Protein technology, developed over several years at the Molecular Biosensor and Imaging Laboratory at Carnegie Mellon University and exclusively licensed to Sharp Edge Labs, will be adapted to the detection of proteins, posttranslational modifications, and protein complexes in a simple add-and- read fluorescence format. The technical innovation involves the creation of a new category of proximity assay with application to the study of both proteins and nucleic acids. The near term goal of the project is the creation of reagents for the multiplexed detection of important tumor markers including p53, EGFR and Her1,2,3. The long-term goal of the project is the creation of a general platform that can be used to create multiplexed assays for the detection of proteins, protein activation profiles, protein-protein interaction profiles, epigenetic modification of individual genes, gene mutations, and compartmental trafficking of proteins. We will test the hypothesis that FAP/Fluorogen binding can be used to create a proximity-ligation type assay platform. We will examine the spatial and energetic requirements of the FAP- Fluorogen interaction to reduce background signal while maximizing detection sensitivity. We will apply the resulting reagents to competitors for EGFR receptor activation. Phase II objectives will address issues of sensitivity, robustness to interference, and applicability across protein and nucleic acid applications. The research reagents market is valued at over 3B annually, with the greatest growth coming from new detectionmethods like those proposed here. We expect our first applications to be in this market, as is the case been for other applications of the FAP technology. We plan to partner with an existing market player for distribution, with the company manufacturing the key detection reagents. The proposed assay format has significant advantages for the companion diagnostics/personalized medicine market, both because of the biology that can be accessed, and because the detection method is simple, creating the possibility of point-of-care instrumentation. This is a large ( 30B) and rapidly expanding market and the company is already establishing connections with distribution partners in this area. PUBLIC HEALTH RELEVANCE: The ability to detect biological molecules, their activation states and their interaction partners is fundamental to understanding biology, to creating treatments for disease, and to developing diagnostics to predict disease and guide treatment. Having exclusively licensed technology developed over many years at the Molecular Biosensor and Imaging Center at Carnegie Mellon University, [1-5] Sharp Edge Labs is developing an innovative method for detecting biological molecules with the ability to elucidate complex biology in a format that can be deployed at the point of patient treatment, improving care and medical treatment outcomes.