Multiplexed Profiling of Extracellular Vesicles for Biomarker Development.

Extracellular vesicles (EVs) are cell-derived membranous particles that play a crucial role in molecular trafficking, intercellular transport and the egress of unwanted proteins. They have been implicated in many diseases including cancer and neurodegeneration. EVs are detected in all bodily fluids, and their protein and nucleic acid content offers a means of assessing the status of the cells from which they originated. As such, they provide opportunities in biomarker discovery for diagnosis, prognosis or the stratification of diseases as well as an objective monitoring of therapies. The simultaneous assaying of multiple EV-derived markers will be required for an impactful practical application, and multiplexing platforms have evolved with the potential to achieve this. Herein, we provide a comprehensive overview of the currently available multiplexing platforms for EV analysis, with a primary focus on miniaturized and integrated devices that offer potential step changes in analytical power, throughput and consistency. Highlights: 1 Extracellular vesicle (EV) multiplexing involves chemical, physical spatial, biological, or nanoparticle-based strategies.2 Multiplexing in EV biomarker development includes high-throughput screening as well as point-of-care testing platforms which to date have been applied mainly to EV surface proteins or internal cargo miRNAs.3 Multiplexed measurements at single-EV resolution are likely to revolutionize the applicability of EV analytes as biomarkers in complex and heterogeneous diseases.