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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV170062	1/1	Homo sapiens	Blood plasma	(d)(U)C ExoQuick IAF	Mustapic M	2017	50%
Study summary Full title Plasma Extracellular Vesicles Enriched for Neuronal Origin: A Potential Window into Brain Pathologic Processes. All authors Mustapic M, Eitan E, Werner JK, Berkowitz ST, Lazaropoulos MP, Tran J, Goetzl EJ, Kapogiannis D Journal J Cell Sci Abstract Our team has been a pioneer in harvesting extracellular vesicles (EVs) enriched for neuronal origin (show more...)Our team has been a pioneer in harvesting extracellular vesicles (EVs) enriched for neuronal origin from peripheral blood and using them as a biomarker discovery platform for neurological disorders. This methodology has demonstrated excellent diagnostic and predictive performance for Alzheimer's and other neurodegenerative diseases in multiple studies, providing a strong proof of concept for this approach. Here, we describe our methodology in detail and offer further evidence that isolated EVs are enriched for neuronal origin. In addition, we present evidence that EVs enriched for neuronal origin represent a more sensitive and accurate base for biomarkers than plasma, serum, or non-enriched total plasma EVs. Finally, we proceed to investigate the protein content of EVs enriched for neuronal origin and compare it with other relevant enriched and non-enriched populations of plasma EVs. Neuronal-origin enriched plasma EVs contain higher levels of signaling molecules of great interest for cellular metabolism, survival, and repair, which may be useful as biomarkers and to follow response to therapeutic interventions in a mechanism-specific manner. (hide) EV-METRIC 50% (82nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Blood plasma Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C ExoQuick IAF Protein markers EV: TSG101/ MAP2/ gamma-enolase/ L1CAMandCD81/ L1CAM/ pTau23/ CD9/ tubulin-betaIII non-EV: None Proteomics no Show all info Study aim Biomarker, Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Blood plasma Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Commercial kit ExoQuick Immunoaffinity capture Selected surface protein(s) L1CAM Other Name other separation method ExoQuick Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins CD9, L1CAM, MAP2, gamma-enolase, pTau23, tubulin-betaIII ELISA Other 1 Proteome Profiler Human Phospho-Mitogen-activated Protein Kinase (MAPK) Antibody Array (Catalog # ARY002B), Human Kidney Biomarker Antibody Array (Catalog # ARY019), Human Phospho-Kinase Antibody Arra Characterization: Lipid analysis No Characterization: Particle analysis PMID previous EV particle analysis Nanoparticle tracking analysis Extra particle analysis NTA Report type Size range/distribution EV concentration Yes EM EM-type Immune-EM EM protein L1CAM and CD81 Image type Close-up, Wide-field

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EV-TRACK ID	EV170062
species	Homo sapiens
sample type	Blood plasma
condition	Control condition
separation protocol	(d)(U)C ExoQuick IAF
Exp. nr.	1
EV-METRIC %	50