Show simple item record Osteikoetxea, Xabier Balogh, Andrea Szabó-Taylor, Katalin Németh, Andrea Szabó, Géza Tamás Pálóczi, Krisztina Sódar, Barbara Kittel, Ágnes György, Bence Pállinger, Éva Matkó, János Buzás, Edit Irén 2015-06-24T08:57:11Z 2015-06-24T08:57:11Z 2015
dc.identifier.citation pagination=e0121184;journalVolume=10;journalIssueNumber=3;journalTitle=PLOS ONE; hu
dc.identifier.uri doi:10.1371/journal.pone.0121184
dc.description.abstract In recent years the study of extracellular vesicles has gathered much scientific and clinical interest. As the field is expanding, it is becoming clear that better methods for characterization and quantification of extracellular vesicles as well as better standards to compare studies are warranted. The goal of the present work was to find improved parameters to characterize extracellular vesicle preparations. Here we introduce a simple 96 well plate-based total lipid assay for determination of lipid content and protein to lipid ratios of extracellular vesicle preparations from various myeloid and lymphoid cell lines as well as blood plasma. These preparations included apoptotic bodies, microvesicles/microparticles, and exosomes isolated by size-based fractionation. We also investigated lipid bilayer order of extracellular vesicle subpopulations using Di-4-ANEPPDHQ lipid probe, and lipid composition using affinity reagents to clustered cholesterol (monoclonal anti-cholesterol antibody) and ganglioside GM1 (cholera toxin subunit B). We have consistently found different protein to lipid ratios characteristic for the investigated extracellular vesicle subpopulations which were substantially altered in the case of vesicular damage or protein contamination. Spectral ratiometric imaging and flow cytometric analysis also revealed marked differences between the various vesicle populations in their lipid order and their clustered membrane cholesterol and GM1 content. Our study introduces for the first time a simple and readily available lipid assay to complement the widely used protein assays in order to better characterize extracellular vesicle preparations. Besides differentiating extracellular vesicle subpopulations, the novel parameters introduced in this work (protein to lipid ratio, lipid bilayer order, and lipid composition), may prove useful for quality control of extracellular vesicle related basic and clinical studies. hu
dc.relation.ispartof urn:issn:1932-6203
dc.title Improved Characterization of EV Preparations Based on Protein to Lipid Ratio and Lipid Properties. hu
dc.type Journal Article hu 2015-06-24T08:46:31Z
dc.language.rfc3066 en hu
dc.identifier.mtmt 2871422
dc.identifier.wos 000351987300224
dc.identifier.pubmed 25798862
dc.contributor.department SE/AOK/I/Genetikai, Sejt- és Immunbiológiai Intézet
dc.contributor.department ELTE/TTK/Bio_I/Immunológiai Tanszék
dc.contributor.institution Semmelweis Egyetem
dc.contributor.institution Eötvös Loránd Tudományegyetem

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