Tools and Methods for Producing High Quality Functional Protein Microarrays for Biomarker Discovery

Abstract ProjectAdministrative Supplements to projectTools and Methods for Producing HighQuality Functional Protein Microarrays for Biomarker Discovery INanoBio is developing ProCap system capable of semi automated production of functional protein arrayswhich can be phosphorylatedde phosphorylated by kinasesphosphatases in high throughput formatin addition to other PTM modificationsAs described in our original SBIR project research planthe ultimate goal is to use ProCap system to produce NAPPA IPC protein arrays on biosensor arraysfor real time detection of proteinproteinproteinenzyme interactions in high throughput formatINanoBio aims to expand ProCap system to integrate protein microarrays with biosensorsthat will boost ADRD research on identifying disease signaling pathwaysdiscovering biomarkers for diagnostics and importantly for drug discovery screeningWe have developed an advanced field effect transistorFETnano sensor technology that can electronically detect Tau protein phosphorylation and de phosphorylation with ultra high sensitivity in real timeyielding kinetic readoutsTau protein interaction with kinases and phosphatases plays a critical role in its hyper phosphorylation that results in aggregation and formation of neruo toxic neurofibrillary tanglesa hallmark of Alzheimerandapos s diseaseUsing supplemental fundingthe project scope will be expanded to test methods for using ProCap system to produce and immobilize protein arrays comprising Tau kinasesphosphatasesfollowed by detecting Tau phosphorylationde phosphorylation using FET nano sensor devices Project Narrative ProjectAdministrative Supplements to projectTools and Methods for Producing HighQuality Functional Protein Microarrays for Biomarker Discovery Commercial platforms capable of producing high quality microarrays of thousands of functional proteins are currently not availableSuch versatile proteome on chip capability will boost proteomics researchand facilitate diagnostics and drug discovery researchINanoBio is developing a proprietary platform termed ProCap that will enable such proteome on chip capabilityThe utility of this platform will be expanded by integrating with novel FDEC FET nanowire sensorsand applying this to detect phosphorylation status of Tau protein that forms neuro toxic tangles in ADRDIn the proposed studywe will demonstrate ProCap system production of ADRD related Tau kinasesphosphatases and detecting Tau phosphorylationdephosphorylation using INanoBio s FDEC FET nano sensors