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You searched for: EV200148 (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
EV200148 1/4 Homo sapiens Blood plasma "(d)(U)C
Filtration
SEC (non-commercial)
UF" 		Menon, Ramkumar 2019 44%

Study summary

Full title
Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated With Term and Preterm Birth
All authors
Ramkumar Menon, Christopher Luke Dixon, Samantha Sheller-Miller, Stephen J Fortunato, George R Saade, Carlos Palma, Andrew Lai, Dominic Guanzon, Carlos Salomon
Journal
Endocrinology
Abstract
Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study (show more...)Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study determined changes in maternal plasma exosome proteomics contents in term and preterm births. Maternal plasma (MP) samples were collected from group 1: term not in labor (TNIL, n = 13); group 2: term in labor (TL, n = 11); group 3: preterm premature rupture of membranes (pPROM, n = 8); and group 4: preterm birth (PTB, n = 13). Exosomes isolated from plasma by differential density centrifugation followed by size exclusion chromatography were characterized by morphology (electron microscopy), quantity and size (nanoparticle tracking analysis), and markers (western blot). A quantitative, information-independent acquisition [sequential windowed acquisition of all theoretical mass spectra (SWATH-MS)] approach was used to determine the protein profile in exosomes. Ingenuity Pathway Analysis determined pathways associated with the protein profile identified in exosomes. MP exosomes were spherical, had a mean diameter of 120 nm, and were positive for exosomal proteins CD63 and TSG101 irrespective of pregnancy status. No distinct changes in exosome quantities were seen in maternal circulation across the groups. SWATH-MS identified 72 statistically significant proteins across the groups studied. Bioinformatics analysis showed the proteins within the exosomes in TNIL, TL, pPROM, and PTB target pathways mainly associated with inflammatory and metabolic signals. Exosomal data suggest that homeostatic imbalances, specifically inflammatory and endocrine signaling, might disrupt pregnancy maintenance resulting in labor-related changes both at term and preterm. Reflection of physiologic changes in exosomes is suggestive of its usefulness as biomarkers and cellular function indicators. (hide)
EV-METRIC
44% (77th 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
"Pregnant; Term, not in labor"
Focus vesicles
exosome
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
Size exclusion chromatography (non-commercial)
Ultrafiltration"
Protein markers
EV: "TSG101/ PLAP/ CD63/ CD9"
non-EV: Grp94
Proteomics
yes
Show all info
Study aim
Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 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
T-8100
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
10
Wash: time (min)
120
Wash: Rotor Type
T-8100
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Not specified
Size-exclusion chromatography
Total column volume (mL)
Not specified
Sample volume/column (mL)
0.5
Resin type
Sepharose CL-2B
Other
Name other separation method
"(Differential) (ultra)centrifugation
Other
Name other separation method
Size exclusion chromatography (non-commercial)
Other
Name other separation method
Ultrafiltration"
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Detected EV-associated proteins
"CD9/ CD63/ TSG101"
Not detected contaminants
Grp94
Proteomics database
No
Fluorescent NTA
Antibody details provided?
No
Detected EV-associated proteins
"PLAP/ CD63"
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
134
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up
EV200148 2/4 Homo sapiens Blood plasma "(d)(U)C
Filtration
SEC (non-commercial)
UF" 		Menon, Ramkumar 2019 33%

Study summary

Full title
Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated With Term and Preterm Birth
All authors
Ramkumar Menon, Christopher Luke Dixon, Samantha Sheller-Miller, Stephen J Fortunato, George R Saade, Carlos Palma, Andrew Lai, Dominic Guanzon, Carlos Salomon
Journal
Endocrinology
Abstract
Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study (show more...)Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study determined changes in maternal plasma exosome proteomics contents in term and preterm births. Maternal plasma (MP) samples were collected from group 1: term not in labor (TNIL, n = 13); group 2: term in labor (TL, n = 11); group 3: preterm premature rupture of membranes (pPROM, n = 8); and group 4: preterm birth (PTB, n = 13). Exosomes isolated from plasma by differential density centrifugation followed by size exclusion chromatography were characterized by morphology (electron microscopy), quantity and size (nanoparticle tracking analysis), and markers (western blot). A quantitative, information-independent acquisition [sequential windowed acquisition of all theoretical mass spectra (SWATH-MS)] approach was used to determine the protein profile in exosomes. Ingenuity Pathway Analysis determined pathways associated with the protein profile identified in exosomes. MP exosomes were spherical, had a mean diameter of 120 nm, and were positive for exosomal proteins CD63 and TSG101 irrespective of pregnancy status. No distinct changes in exosome quantities were seen in maternal circulation across the groups. SWATH-MS identified 72 statistically significant proteins across the groups studied. Bioinformatics analysis showed the proteins within the exosomes in TNIL, TL, pPROM, and PTB target pathways mainly associated with inflammatory and metabolic signals. Exosomal data suggest that homeostatic imbalances, specifically inflammatory and endocrine signaling, might disrupt pregnancy maintenance resulting in labor-related changes both at term and preterm. Reflection of physiologic changes in exosomes is suggestive of its usefulness as biomarkers and cellular function indicators. (hide)
EV-METRIC
33% (65th 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
"Pregnant; Term, in labor"
Focus vesicles
exosome
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
Size exclusion chromatography (non-commercial)
Ultrafiltration"
Protein markers
EV: "TSG101/ PLAP/ CD63/ CD9"
non-EV: Grp94
Proteomics
yes
Show all info
Study aim
Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 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
T-8100
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
10
Wash: time (min)
120
Wash: Rotor Type
T-8100
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Not specified
Size-exclusion chromatography
Total column volume (mL)
Not specified
Sample volume/column (mL)
0.5
Resin type
Sepharose CL-2B
Other
Name other separation method
"(Differential) (ultra)centrifugation
Other
Name other separation method
Size exclusion chromatography (non-commercial)
Other
Name other separation method
Ultrafiltration"
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Detected EV-associated proteins
"CD9/ CD63/ TSG101"
Not detected contaminants
Grp94
Proteomics database
No
Fluorescent NTA
Antibody details provided?
No
Detected EV-associated proteins
"PLAP/ CD63"
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
113
EV concentration
Yes
EV200148 3/4 Homo sapiens Blood plasma "(d)(U)C
Filtration
SEC (non-commercial)
UF" 		Menon, Ramkumar 2019 33%

Study summary

Full title
Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated With Term and Preterm Birth
All authors
Ramkumar Menon, Christopher Luke Dixon, Samantha Sheller-Miller, Stephen J Fortunato, George R Saade, Carlos Palma, Andrew Lai, Dominic Guanzon, Carlos Salomon
Journal
Endocrinology
Abstract
Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study (show more...)Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study determined changes in maternal plasma exosome proteomics contents in term and preterm births. Maternal plasma (MP) samples were collected from group 1: term not in labor (TNIL, n = 13); group 2: term in labor (TL, n = 11); group 3: preterm premature rupture of membranes (pPROM, n = 8); and group 4: preterm birth (PTB, n = 13). Exosomes isolated from plasma by differential density centrifugation followed by size exclusion chromatography were characterized by morphology (electron microscopy), quantity and size (nanoparticle tracking analysis), and markers (western blot). A quantitative, information-independent acquisition [sequential windowed acquisition of all theoretical mass spectra (SWATH-MS)] approach was used to determine the protein profile in exosomes. Ingenuity Pathway Analysis determined pathways associated with the protein profile identified in exosomes. MP exosomes were spherical, had a mean diameter of 120 nm, and were positive for exosomal proteins CD63 and TSG101 irrespective of pregnancy status. No distinct changes in exosome quantities were seen in maternal circulation across the groups. SWATH-MS identified 72 statistically significant proteins across the groups studied. Bioinformatics analysis showed the proteins within the exosomes in TNIL, TL, pPROM, and PTB target pathways mainly associated with inflammatory and metabolic signals. Exosomal data suggest that homeostatic imbalances, specifically inflammatory and endocrine signaling, might disrupt pregnancy maintenance resulting in labor-related changes both at term and preterm. Reflection of physiologic changes in exosomes is suggestive of its usefulness as biomarkers and cellular function indicators. (hide)
EV-METRIC
33% (65th 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
"Pregnant; Preterm, premature rupture of membranes"
Focus vesicles
exosome
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
Size exclusion chromatography (non-commercial)
Ultrafiltration"
Protein markers
EV: "TSG101/ PLAP/ CD63/ CD9"
non-EV: Grp94
Proteomics
yes
Show all info
Study aim
Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 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
T-8100
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
10
Wash: time (min)
120
Wash: Rotor Type
T-8100
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Not specified
Size-exclusion chromatography
Total column volume (mL)
Not specified
Sample volume/column (mL)
0.5
Resin type
Sepharose CL-2B
Other
Name other separation method
"(Differential) (ultra)centrifugation
Other
Name other separation method
Size exclusion chromatography (non-commercial)
Other
Name other separation method
Ultrafiltration"
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Detected EV-associated proteins
"CD9/ CD63/ TSG101"
Not detected contaminants
Grp94
Proteomics database
No
Fluorescent NTA
Antibody details provided?
No
Detected EV-associated proteins
"PLAP/ CD63"
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
120
EV concentration
Yes
EV200148 4/4 Homo sapiens Blood plasma "(d)(U)C
Filtration
SEC (non-commercial)
UF" 		Menon, Ramkumar 2019 33%

Study summary

Full title
Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated With Term and Preterm Birth
All authors
Ramkumar Menon, Christopher Luke Dixon, Samantha Sheller-Miller, Stephen J Fortunato, George R Saade, Carlos Palma, Andrew Lai, Dominic Guanzon, Carlos Salomon
Journal
Endocrinology
Abstract
Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study (show more...)Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study determined changes in maternal plasma exosome proteomics contents in term and preterm births. Maternal plasma (MP) samples were collected from group 1: term not in labor (TNIL, n = 13); group 2: term in labor (TL, n = 11); group 3: preterm premature rupture of membranes (pPROM, n = 8); and group 4: preterm birth (PTB, n = 13). Exosomes isolated from plasma by differential density centrifugation followed by size exclusion chromatography were characterized by morphology (electron microscopy), quantity and size (nanoparticle tracking analysis), and markers (western blot). A quantitative, information-independent acquisition [sequential windowed acquisition of all theoretical mass spectra (SWATH-MS)] approach was used to determine the protein profile in exosomes. Ingenuity Pathway Analysis determined pathways associated with the protein profile identified in exosomes. MP exosomes were spherical, had a mean diameter of 120 nm, and were positive for exosomal proteins CD63 and TSG101 irrespective of pregnancy status. No distinct changes in exosome quantities were seen in maternal circulation across the groups. SWATH-MS identified 72 statistically significant proteins across the groups studied. Bioinformatics analysis showed the proteins within the exosomes in TNIL, TL, pPROM, and PTB target pathways mainly associated with inflammatory and metabolic signals. Exosomal data suggest that homeostatic imbalances, specifically inflammatory and endocrine signaling, might disrupt pregnancy maintenance resulting in labor-related changes both at term and preterm. Reflection of physiologic changes in exosomes is suggestive of its usefulness as biomarkers and cellular function indicators. (hide)
EV-METRIC
33% (65th 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
"Pregnant; Preterm birth"
Focus vesicles
exosome
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
Size exclusion chromatography (non-commercial)
Ultrafiltration"
Protein markers
EV: "TSG101/ PLAP/ CD63/ CD9"
non-EV: Grp94
Proteomics
yes
Show all info
Study aim
Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 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
T-8100
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
10
Wash: time (min)
120
Wash: Rotor Type
T-8100
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Not specified
Size-exclusion chromatography
Total column volume (mL)
Not specified
Sample volume/column (mL)
0.5
Resin type
Sepharose CL-2B
Other
Name other separation method
"(Differential) (ultra)centrifugation
Other
Name other separation method
Size exclusion chromatography (non-commercial)
Other
Name other separation method
Ultrafiltration"
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Detected EV-associated proteins
"CD9/ CD63/ TSG101"
Not detected contaminants
Grp94
Proteomics database
No
Fluorescent NTA
Antibody details provided?
No
Detected EV-associated proteins
"PLAP/ CD63"
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
126
EV concentration
Yes
1 - 4 of 4
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV200148
species
Homo sapiens
sample type
Blood plasma
condition
Pregnant
Term
not in labor
Pregnant
Term
in labor
Pregnant
Preterm
premature rupture of membranes
Pregnant
Preterm birth
separation protocol
dUC
Filtration
Size exclusion chromatography (non-commercial)
Ultrafiltration
dUC
Filtration
Size exclusion chromatography (non-commercial)
Ultrafiltration
dUC
Filtration
Size exclusion chromatography (non-commercial)
Ultrafiltration
dUC
Filtration
Size exclusion chromatography (non-commercial)
Ultrafiltration
Exp. nr.
1
2
3
4
EV-METRIC %
44
33
33
33