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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230654	1/1	Mycobacterium tuberculosis	H37Rv	(d)(U)C DC DG Filtration UF	Lee J	2015	63%
Study summary Full title Proteomic analysis of extracellular vesicles derived from Mycobacterium tuberculosis. All authors Lee J, Kim SH, Choi DS, Lee JS, Kim DK, Go G, Park SM, Kim SH, Shin JH, Chang CL, Gho YS Journal Proteomics Abstract The release of extracellular vesicles, also known as outer membrane vesicles, membrane vesicles, exo (show more...)The release of extracellular vesicles, also known as outer membrane vesicles, membrane vesicles, exosomes, and microvesicles, is an evolutionarily conserved phenomenon from bacteria to eukaryotes. It has been reported that Mycobacterium tuberculosis releases extracellular vesicles harboring immunologically active molecules, and these extracellular vesicles have been suggested to be applicable in vaccine development and biomarker discovery. However, the comprehensive proteomic analysis has not been performed for M. tuberculosis extracellular vesicles. In this study, we identified a total of 287 vesicular proteins by four LC-MS/MS analyses with high confidence. In addition, we identified several vesicular proteins associated with the virulence of M. tuberculosis. This comprehensive proteome profile will help elucidate the pathogenic mechanism of M. tuberculosis. The data have been deposited to the ProteomeXchange with identifier PXD001160 (http://proteomecentral.proteomexchange.org/dataset/PXD001160). (hide) EV-METRIC 63% (93rd 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 Density cushion Density gradient Filtration Ultrafiltration Protein markers EV: LprG non-EV: None Proteomics yes EV density (g/ml) 1,17 Show all info Study aim Identification of content (omics approaches) Sample Species Mycobacterium tuberculosis Sample Type Cell culture supernatant EV-producing cells H37Rv EV-harvesting Medium serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Pelleting performed No Density gradient Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 10% Highest density fraction 60% Total gradient volume, incl. sample (mL) 10,3 Sample volume (mL) 4,8 Orientation Bottom-up Speed (g) 200000 Duration (min) 120 Fraction volume (mL) 1 Fraction processing None Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Density cushion Density medium Sucrose Cushion volume 1,5 Density of the cushion 0.8-2.0 M Centrifugation time 120 Centrifugation speed 100000 Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Detected EV-associated proteins LprG Proteomics database ProteomeXchange Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 40-50 EV concentration Yes Particle yield Total particle count EM EM-type Transmission-EM Image type Wide-field Report size (nm) 20-250

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EV-TRACK ID	EV230654
species	Mycobacterium tuberculosis
sample type	Cell culture
cell type	H37Rv
condition	Control condition
separation protocol	dUC/ DC/ Density gradient/ Filtration/ Ultrafiltration
Exp. nr.	1
EV-METRIC %	63