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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV190083	1/2	Pectobacterium betavasculorum	IFB5271	(d)(U)C	Piotrowska M	2020	71%
Study summary Full title Capillary zone electrophoresis of bacterial extracellular vesicles: A proof of concept. All authors Piotrowska M, Ciura K, Zalewska M, Dawid M, Correia B, Sawicka P, Lewczuk B, Kasprzyk J, Sola L, Piekoszewski W, Wielgomas B, Waleron K, Dziomba S Journal J Chromatogr A Abstract The extracellular vesicles (EVs) released by plant pathogens of the Pectobacterium genus were invest (show more...)The extracellular vesicles (EVs) released by plant pathogens of the Pectobacterium genus were investigated. The isolates were obtained using differential centrifugation followed by filtration and were characterized in terms of total protein content and particle size distribution. The transmission electron microscopy (TEM) analysis revealed the presence of two morphologically differentiated subpopulations of vesicles in the obtained isolates. The proteomic analysis using matrix-assisted laser desorption ionization mass spectrometry with time of flight detector (MALDI-TOF/TOF-MS) enabled to identify 62 proteomic markers commonly found in EVs of Gram-negative rods from the Enterobacteriaceae family. Capillary electrophoresis (CE) was proposed as a novel tool for the characterization of EVs. The method allowed for automated and fast (<15 min per sample) separation of vesicles from macromolecular aggregates with low sample consumption (about 10 nL per analysis). The approach required simple background electrolyte (BGE) composed of 50 mM BTP and 75 mM glycine (pH 9.5) and standard UV detection. The report presents a new opportunity for quality control of samples containing EVs. (hide) EV-METRIC 71% (96th 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 Cell culture supernatant 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 (Differential) (ultra)centrifugation Protein markers EV: None non-EV: GroEL Proteomics yes Show all info Study aim New methodological development/Identification of content (omics approaches) Sample Species Pectobacterium betavasculorum Sample Type Cell culture supernatant EV-producing cells IFB5271 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type SW 28 Pelleting: speed (g) 85000 Characterization: Protein analysis Protein Concentration Method Lowry-based assay Protein Yield (µg) Not reported Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 514 EM EM-type Transmission-EM Image type Close-up, Wide-field Other particle analysis name(1) Capillary electrophoresis EV-concentration No

	EV190083	2/2	Pectobacterium betavasculorum	IFB5271	(d)(U)C Filtration	Piotrowska M	2020	71%
Study summary Full title Capillary zone electrophoresis of bacterial extracellular vesicles: A proof of concept. All authors Piotrowska M, Ciura K, Zalewska M, Dawid M, Correia B, Sawicka P, Lewczuk B, Kasprzyk J, Sola L, Piekoszewski W, Wielgomas B, Waleron K, Dziomba S Journal J Chromatogr A Abstract The extracellular vesicles (EVs) released by plant pathogens of the Pectobacterium genus were invest (show more...)The extracellular vesicles (EVs) released by plant pathogens of the Pectobacterium genus were investigated. The isolates were obtained using differential centrifugation followed by filtration and were characterized in terms of total protein content and particle size distribution. The transmission electron microscopy (TEM) analysis revealed the presence of two morphologically differentiated subpopulations of vesicles in the obtained isolates. The proteomic analysis using matrix-assisted laser desorption ionization mass spectrometry with time of flight detector (MALDI-TOF/TOF-MS) enabled to identify 62 proteomic markers commonly found in EVs of Gram-negative rods from the Enterobacteriaceae family. Capillary electrophoresis (CE) was proposed as a novel tool for the characterization of EVs. The method allowed for automated and fast (<15 min per sample) separation of vesicles from macromolecular aggregates with low sample consumption (about 10 nL per analysis). The approach required simple background electrolyte (BGE) composed of 50 mM BTP and 75 mM glycine (pH 9.5) and standard UV detection. The report presents a new opportunity for quality control of samples containing EVs. (hide) EV-METRIC 71% (96th 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 Cell culture supernatant 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 (Differential) (ultra)centrifugation Filtration Protein markers EV: None non-EV: GroEL Proteomics yes Show all info Study aim New methodological development/Identification of content (omics approaches) Sample Species Pectobacterium betavasculorum Sample Type Cell culture supernatant EV-producing cells IFB5271 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: rotor type SW 28 Pelleting: speed (g) 85000 Filtration steps 0.2 or 0.22 µm Characterization: Protein analysis Protein Concentration Method Lowry-based assay Protein Yield (µg) Not reported Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 184 EM EM-type Transmission-EM Image type Close-up, Wide-field Other particle analysis name(1) Capillary electrophoresis EV-concentration No

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EV-TRACK ID	EV190083
species	Pectobacterium betavasculorum
sample type	Cell culture
cell type	IFB5271
condition	Control condition
separation protocol	dUC	dUC/ Filtration
Exp. nr.	1	2
EV-METRIC %	71	71