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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV220081	2/2	Homo sapiens	Peritoneal dialysis efflux	(d)(U)C SEC (non-commercial) UF Filtration	Carreras-Planella L	2017	43%
Study summary Full title Characterization and proteomic profile of extracellular vesicles from peritoneal dialysis efflux. All authors Carreras-Planella L, Soler-Majoral J, Rubio-Esteve C, Lozano-Ramos SI, Franquesa M, Bonet J, Troya-Saborido MI, Borràs FE Journal PLoS One Abstract Peritoneal Dialysis (PD) is considered the best option for a cost-effective mid-term dialysis in pat (show more...)Peritoneal Dialysis (PD) is considered the best option for a cost-effective mid-term dialysis in patients with Chronic Renal Failure. However, functional failure of the peritoneal membrane (PM) force many patients to stop PD treatment and start haemodialysis. Currently, PM functionality is monitored by the peritoneal equilibration test, a tedious technique that often show changes when the membrane damage is advanced. As in other pathologies, the identification and characterization of extracellular vesicles (EVs) in the peritoneal dialysis efflux (PDE) may represent a non-invasive alternative to identify biomarkers of membrane failure. Using size-exclusion chromatography, we isolated EVs from PDE in a group of patients. Vesicles were characterized by the presence of tetraspanin markers, nanoparticle tracking analysis profile, cryo-electron microscopy and mass spectrometry. Here, we report the isolation and characterization of PDE-EVs. Based on mass spectrometry, we have found a set of well-conserved proteins among patients. Interestingly, the peptide profile also revealed remarkable changes between newly enrolled and longer-treated PD patients. These results are the first step to the identification of PDE-EVs based new markers of PM damage, which could support clinicians in their decision-making in a non-invasive manner. (hide) EV-METRIC 43% (75th 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 Peritoneal dialysis efflux Sample origin >18 months on peritoneal dialysis 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 (Differential) (ultra)centrifugation Size-exclusion chromatography (non-commercial) Ultrafiltration Filtration Protein markers EV: CD63/ CD9 non-EV: None Proteomics yes Show all info Study aim Biomarker/Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Peritoneal dialysis efflux Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Regenerated cellulose Size-exclusion chromatography Total column volume (mL) 12 Sample volume/column (mL) 0.8-2 Characterization: Protein analysis Protein Concentration Method Bradford Flow cytometry aspecific beads Antibody details provided? No Detected EV-associated proteins CD9/ CD63 Flow cytometry specific beads Antibody details provided? Yes Antibody dilution provided? No Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 100-200 EV concentration Yes EM EM-type Cryo-EM Image type Wide-field

	EV220081	1/2	Homo sapiens	Peritoneal dialysis efflux	(d)(U)C SEC (non-commercial) UF Filtration	Carreras-Planella L	2017	29%
Study summary Full title Characterization and proteomic profile of extracellular vesicles from peritoneal dialysis efflux. All authors Carreras-Planella L, Soler-Majoral J, Rubio-Esteve C, Lozano-Ramos SI, Franquesa M, Bonet J, Troya-Saborido MI, Borràs FE Journal PLoS One Abstract Peritoneal Dialysis (PD) is considered the best option for a cost-effective mid-term dialysis in pat (show more...)Peritoneal Dialysis (PD) is considered the best option for a cost-effective mid-term dialysis in patients with Chronic Renal Failure. However, functional failure of the peritoneal membrane (PM) force many patients to stop PD treatment and start haemodialysis. Currently, PM functionality is monitored by the peritoneal equilibration test, a tedious technique that often show changes when the membrane damage is advanced. As in other pathologies, the identification and characterization of extracellular vesicles (EVs) in the peritoneal dialysis efflux (PDE) may represent a non-invasive alternative to identify biomarkers of membrane failure. Using size-exclusion chromatography, we isolated EVs from PDE in a group of patients. Vesicles were characterized by the presence of tetraspanin markers, nanoparticle tracking analysis profile, cryo-electron microscopy and mass spectrometry. Here, we report the isolation and characterization of PDE-EVs. Based on mass spectrometry, we have found a set of well-conserved proteins among patients. Interestingly, the peptide profile also revealed remarkable changes between newly enrolled and longer-treated PD patients. These results are the first step to the identification of PDE-EVs based new markers of PM damage, which could support clinicians in their decision-making in a non-invasive manner. (hide) EV-METRIC 29% (25th 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 Peritoneal dialysis efflux Sample origin < 10 months on peritoneal dialysis 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 (Differential) (ultra)centrifugation Size-exclusion chromatography (non-commercial) Ultrafiltration Filtration Protein markers EV: CD63/ CD9 non-EV: None Proteomics yes Show all info Study aim Biomarker/Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Peritoneal dialysis efflux Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Regenerated cellulose Size-exclusion chromatography Total column volume (mL) 12 Sample volume/column (mL) 0.8-2 Characterization: Protein analysis Protein Concentration Method Bradford Flow cytometry aspecific beads Antibody details provided? No Detected EV-associated proteins CD9/ CD63 Flow cytometry specific beads Antibody details provided? Yes Antibody dilution provided? No Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA EV concentration Yes

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EV-TRACK ID	EV220081
species	Homo sapiens
sample type	Peritoneal dialysis efflux
condition	>18 months on peritoneal dialysis	< 10 months on peritoneal dialysis
separation protocol	dUC/ Size-exclusion chromatography (non-commercial)/ Ultrafiltration/ Filtration	dUC/ Size-exclusion chromatography (non-commercial)/ Ultrafiltration/ Filtration
Exp. nr.	2	1
EV-METRIC %	43	29