Abstract
Myeloperoxidase (MPO) is a circulating cardiovascular disease (CVD) biomarker used to estimate clinical risk and patient prognosis. Current enzyme-linked immunosorbent assays (ELISA) for MPO concentration are costly and time-intensive. Here we report a novel bioluminescence assay, designated MPO activity on a polymer surface (MAPS), for measuring MPO activity in human plasma samples using the bioluminescent substrate L-012. The method delivers a result in under an hour and is resistant to confounding effects from endogenous MPO inhibitors. In a pilot clinical study, we compared MAPS and two clinical ELISAs using 72 plasma samples from cardiac catheterization patients. Results from parallel MAPS and ELISAs were concordant within 2±11 μg l−1 MPO with similar uncertainty and reproducibility. Results between parallel MAPS and ELISA were in better agreement than those between independent ELISAs. MAPS may provide an inexpensive and rapid assay for determining MPO activity in plasma samples from patients with CVD or potentially other immune and inflammatory disorders.
Highlights
Myeloperoxidase (MPO) is a circulating cardiovascular disease (CVD) biomarker used to estimate clinical risk and patient prognosis
Plasma MPO concentration is usually measured by enzyme-linked immunosorbent assay (ELISA)[11], which is costly, time-intensive and typically uses Reactive oxygen species (ROS) generated by immunoconjugate horseradish peroxidase (HRP) instead of directly measuring MPO-derived ROS
We have previously shown that MPO activity can be imaged directly in vivo with luminol, a chemiluminescent compound oxidized by hypochlorous acid[14]
Summary
Myeloperoxidase (MPO) is a circulating cardiovascular disease (CVD) biomarker used to estimate clinical risk and patient prognosis. Current enzyme-linked immunosorbent assays (ELISA) for MPO concentration are costly and time-intensive. Results between parallel MAPS and ELISA were in better agreement than those between independent ELISAs. MAPS may provide an inexpensive and rapid assay for determining MPO activity in plasma samples from patients with CVD or potentially other immune and inflammatory disorders. MPO constitutes 5% of neutrophil dry weight and is concentrated in primary granules[6] On neutrophil activation, these granules fuse to the phagosomal or cell membrane to oxidize biomolecules with hypochlorous acid produced by MPO7. Plasma MPO concentration is usually measured by enzyme-linked immunosorbent assay (ELISA)[11], which is costly, time-intensive and typically uses ROS generated by immunoconjugate horseradish peroxidase (HRP) instead of directly measuring MPO-derived ROS.
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