Single molecule characterization of individual extracellular vesicles from pancreatic cancer.

ABSTRACT

Biofluid-accessible extracellular vesicles (EVs) may represent a new means to improve the sensitivity and specificity of detecting disease. However, current methods to isolate EVs encounter challenges when they are used to select specific populations. Moreover, it has been difficult to comprehensively characterize heterogeneous EV populations at the single vesicle level. Here, we robustly assessed heterogeneous EV populations from cultured cell lines via nanoparticle tracking analysis, proteomics, transcriptomics, transmission electron microscopy, and quantitative single molecule localization microscopy (qSMLM). Using qSMLM, we quantified the size and biomarker content of individual EVs. We applied qSMLM to patient plasma samples and identified a pancreatic cancer-enriched EV population. Our goal is to advance single molecule characterization of EVs for early disease detection. Abbreviations EV Extracellular Vesicle; qSMLM quantitative Single Molecule Localization Microscopy; PDAC Pancreatic Ductal Adenocarcinoma; EGFR epidermal growth factor receptor 1; CA19-9 carbohydrate antigen 19-9; SEC size exclusion chromatography; WGA wheat germ agglutinin; AF647 Alexa Fluor 647; Ab antibody; HPDEC Healthy Pancreatic Ductal Epithelial Cell; TEM Transmission Electron Microscopy.