Project Summary A Chemiluminescent Assay and Device for Monitoring Vascular Endogenous Nitric Oxide This STTR Phase I project aims to establish the feasibility of a novel chemiluminescent assay for the detection of nitric oxide in blood. Phase II will aim to use this assay in a point-of-care device to measure vascular endogenous nitric oxide (VeNO) from a finger-prick of blood for ultimate deployment in a clinical setting. Vascular nitric oxide is fundamental to cardiovascular health and an accurate measure will streamline drug discovery efforts, help monitor patients, and identify patients that are most likely to benefit from therapies that target nitric oxide signaling. This proposal is a collaboration between BioLum Sciences, LLC and the PI’s laboratory at Southern Methodist University. The specific aims include: Aim 1 (BioLum) Optimize and validate a chemiluminescent assay for sensitive and specific nitric oxide measurement in blood Preliminary data has demonstrated the ability of a novel chemiluminescent assay to measure nitric oxide in aqueous buffers. This assay uses the chemiluminescent spiroadamantane 1,2-dioxetane reagent, PNCL, to measure peroxynitrite formed by trapping nitric oxide with superoxide. Using funding from this award, BioLum will optimize this assay for increased sensitivity, characterize its efficacy for measuring nitric oxide in blood samples, and validate the assay with contemporary nitric oxide measurement methods. Aim 2 (SMU) Measure and optimize NO decomposition kinetics under assay conditions Measurement of vascular endogenous nitric oxide in a finger-prick of blood will depend on the decomposition kinetics upon separation from a source of production. These kinetics will be measured using solid-supported nitric oxide donors and stopped-flow spectroscopy techniques. The decomposition rate in aerated buffer in the absence and presence of red blood cells will be determined and assay conditions will be formulated for optimal nitric oxide lifetimes.