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You searched for: EV21008 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Details EV-TRACK ID Experiment nr. Species Sample type Separation protocol First author Year EV-METRIC
EV21008 1/2 Homo sapiens Blood plasma DG
(d)(U)C
Annalisa Radeghieri 2022 89%

Study summary

Full title
All authors
Annalisa Radeghieri, Silvia Alacqua, Andrea Zendrini, Vanessa Previcini, Francesca Todaro, Giuliana Martini, Doris Ricotta, Paolo Bergese
Journal
Journal of Extracellular Biology
Abstract
Antithrombin (AT) is a glycoprotein produced by the liver and a principal antagonist of active clott (show more...)Antithrombin (AT) is a glycoprotein produced by the liver and a principal antagonist of active clotting proteases. A deficit in AT function leads to AT qualitative deficiency, challenging to diagnose. Here we report that active AT may travel physiosorbed on the surface of plasma extracellular vesicles (EVs), contributing to form the “EV-protein corona.” The corona is enriched in specific AT glycoforms, thus suggesting glycosylation to play a key role in AT partitioning between EVs and plasma. Differences in AT glycoform composition of the corona of EVs separated from plasma of healthy and AT qualitative deficiency-affected subjects were also noticed. This suggests deconstructing the plasma into its nanostructured components, as EVs, could suggest novel directions to unravel pathophysiological mechanisms. (hide)
EV-METRIC
89% (99th 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
DG
(d)(U)C
Protein markers
EV: TSG101/ CD63/ CD81/ Adam 10/ Alix/ Antithrombin 3
non-EV: Argonaute2/ Apo A1/ GM130
Proteomics
no
EV density (g/ml)
1.11-1.22
Show all info
Study aim
Function/Biomarker
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
TLA-55
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
10
Lowest density fraction
8%
Highest density fraction
75%
Total gradient volume, incl. sample (mL)
4,8
Sample volume (mL)
1
Orientation
Top-down
Rotor type
MLS-50
Speed (g)
100000
Duration (min)
960
Fraction volume (mL)
0,4
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
120
Pelleting: rotor type
TLA-55
Pelleting: speed (g)
100000
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD63/ Adam 10/ TSG101/ Alix/ CD81
Not detected contaminants
Apo A1/ GM130
Detected EV-associated proteins
CD63/ TSG101
Detected contaminants
Argonaute2
Detected EV-associated proteins
Antithrombin 3
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Atomic force-EM
Image type
Close-up, Wide-field
Report size (nm)
50
EV21008 2/2 Homo sapiens Blood plasma DG
(d)(U)C
Annalisa Radeghieri 2022 50%

Study summary

Full title
All authors
Annalisa Radeghieri, Silvia Alacqua, Andrea Zendrini, Vanessa Previcini, Francesca Todaro, Giuliana Martini, Doris Ricotta, Paolo Bergese
Journal
Journal of Extracellular Biology
Abstract
Antithrombin (AT) is a glycoprotein produced by the liver and a principal antagonist of active clott (show more...)Antithrombin (AT) is a glycoprotein produced by the liver and a principal antagonist of active clotting proteases. A deficit in AT function leads to AT qualitative deficiency, challenging to diagnose. Here we report that active AT may travel physiosorbed on the surface of plasma extracellular vesicles (EVs), contributing to form the “EV-protein corona.” The corona is enriched in specific AT glycoforms, thus suggesting glycosylation to play a key role in AT partitioning between EVs and plasma. Differences in AT glycoform composition of the corona of EVs separated from plasma of healthy and AT qualitative deficiency-affected subjects were also noticed. This suggests deconstructing the plasma into its nanostructured components, as EVs, could suggest novel directions to unravel pathophysiological mechanisms. (hide)
EV-METRIC
50% (83rd 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
Antithrombin 3 deficiency
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
DG
(d)(U)C
Protein markers
EV: Antithrombin 3
non-EV: None
Proteomics
no
EV density (g/ml)
1.11-1.22
Show all info
Study aim
Function/Biomarker
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
TLA-55
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
10
Lowest density fraction
8%
Highest density fraction
75%
Total gradient volume, incl. sample (mL)
4,8
Sample volume (mL)
1
Orientation
Top-down
Rotor type
MLS-50
Speed (g)
100000
Duration (min)
960
Fraction volume (mL)
0,4
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
120
Pelleting: rotor type
TLA-55
Pelleting: speed (g)
100000
Characterization: Protein analysis
Protein Concentration Method
Not determined
Detected EV-associated proteins
Antithrombin 3
Characterization: Lipid analysis
No
Characterization: Particle analysis
None
1 - 2 of 2
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV21008
species
Homo sapiens
sample type
Blood plasma
condition
Control condition
Antithrombin
3 deficiency
separation protocol
DG
(d)(U)C
DG
(d)(U)C
Exp. nr.
1
2
EV-METRIC %
89
50