Abstract A critical bottleneck in the field of proteomics is the inability to simultaneously and cost-effectively evaluate thousands of functional protein interactions at once to obtain kinetic data. Although conventional in-situ protein microarray technology is well developed, it uses fluorescent dye based detection that provides only qualitative or semi-quantitative information. There is currently no technology that offers label-free real-time interaction assays of full-length folded proteins in high throughput format. To address this unmet need, INanoBio is developing novel sensor integrated proteome on chip (SPOC) platform that combines in situ expression and capture of functional proteins on biosensors, to allow instant high throughput analysis of thousands of proteins in real time. In this SBIR phase II project, we propose to develop SPOC platform by integrating in situ produced protein arrays with surface plasmon resonance imaging (SPRi), by building integrated ProSPR instrument that will enable SPRi determination of both qualitative and kinetic information of protein biomolecular interactions in high throughput. We will develop, validate and apply 1000 protein SPOC SPRi sensor chips to study clinically relevant protein- protein interactions and test off-target binding of new antibodies produced in animal models against human proteins.