biostudies-literatureXu YFDivision of Cancer Epidemiology and Genetics, National Cancer InstituteNational Institute of General Medical Sciences of the National Institutes of HealthOklahoma Center for the Advancement of Science and TechnologyNCI NIH HHSPresbyterian Health FoundationPeggy and Charles Stephenson Cancer CenterNIGMS NIH HHSe0259563https://www.ebi.ac.uk/biostudies/studies/S-EPMC8594802biostudies-literatureUnknown16(11)microRNAs (miRNA) in extracellular vesicles (EVs) have been investigated as potential biomarkers for pancreatic ductal adenocarcinoma (PDAC). However, a mixed population of EVs is often obtained using conventional exosome isolation methods for biomarker development. EVs are derived from different cellular processes and present in various sizes, therefore miRNA expression among them is undoubtedly different. We developed a simple protocol utilizing sequential filtration and ultracentrifugation to separate PDAC EVs into three groups, one with an average diameter of more than 220 nm, named operational 3 (OP3); one with average diameters between 100-220 nm, named operational 2 (OP2); and another with average diameters around 100 nm, named operational 1 (OP1)). EVs were isolated from conditionePloS oneIsolation of extra-cellular vesicles in the context of pancreatic adenocarcinomas: Addition of one stringent filtration step improves recovery of specific microRNAs.PMC8594802P20GM103640HR14-147P20 GM103640R03 CA235208CA235208-01Gin ARao CVHannafon BNXu XXu YFBhandari KMorris KTDing WQfalseIsolation of extra-cellular vesicles in the context of pancreatic adenocarcinomas: Addition of one stringent filtration step improves recovery of specific microRNAs.microRNAs (miRNA) in extracellular vesicles (EVs) have been investigated as potential biomarkers for pancreatic ductal adenocarcinoma (PDAC). However, a mixed population of EVs is often obtained using conventional exosome isolation methods for biomarker development. EVs are derived from different cellular processes and present in various sizes, therefore miRNA expression among them is undoubtedly different. We developed a simple protocol utilizing sequential filtration and ultracentrifugation to separate PDAC EVs into three groups, one with an average diameter of more than 220 nm, named operational 3 (OP3); one with average diameters between 100-220 nm, named operational 2 (OP2); and another with average diameters around 100 nm, named operational 1 (OP1)). EVs were isolated from conditione2021-01-01T00:00:00Z20212025-04-22T11:30:51.029Z2022-02-11T12:58:11.678ZS-EPMC85948023478437710.1371/journal.pone.0259563