Search > Results

You searched for: EV210135 (EV-TRACK ID)

Showing 1 - 2 of 2

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
EV210135 1/2 Homo sapiens Cell culture supernatant DG
(d)(U)C
UF
Stremersch, Stephan 2016 78%

Study summary

Full title
All authors
Stephan Stremersch, Monica Marro, Bat-El Pinchasik, Pieter Baatsen, An Hendrix, Stefaan C De Smedt, Pablo Loza-Alvarez, Andre G Skirtach, Koen Raemdonck, Kevin Braeckmans
Journal
Small
Abstract
Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention f (show more...)Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention for the detection of cancer at an early stage. In this study the feasibility of using a surface enhanced Raman spectroscopy (SERS) based method to distinguish between ELVs derived from different cellular origins is evaluated. A gold nanoparticle based shell is deposited on the surface of ELVs derived from cancerous and healthy cells, which enhances the Raman signal while maintaining a colloidal suspension of individual vesicles. This nanocoating allows the recording of SERS spectra from single vesicles. By using partial least squares discriminant analysis on the obtained spectra, vesicles from different origin can be distinguished, even when present in the same mixture. This proof-of-concept study paves the way for noninvasive (cancer) diagnostic tools based on exosomal SERS fingerprinting in combination with multivariate statistical analysis. (hide)
EV-METRIC
78% (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
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
Cell Name
Red blood cells
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.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
DG
(d)(U)C
UF
Protein markers
EV: CD81/ HSP70/ CD63/ ?-actin
non-EV: None
Proteomics
no
EV density (g/ml)
1.14
Show all info
Study aim
Biomarker/Technical analysis comparing/optimizing EV-­related methods/New methodological development
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
Red blood cells
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
12.5%
Highest density fraction
50%
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 55 Ti
Speed (g)
150000
Duration (min)
900
Fraction volume (mL)
0.5
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
150000
Ultra filtration
Cut-off size (kDa)
30
Membrane type
Not specified
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63/ ?-actin/ HSP70/ CD81
Flow cytometry
Hardware adjustments
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
170
Particle yield
Not reported
EM
EM-type
Cryo-­EM
Image type
Wide-field
EV concentration
Not
EV210135 2/2 Mus musculus Cell culture supernatant DG
(d)(U)C
UF
Stremersch, Stephan 2016 78%

Study summary

Full title
All authors
Stephan Stremersch, Monica Marro, Bat-El Pinchasik, Pieter Baatsen, An Hendrix, Stefaan C De Smedt, Pablo Loza-Alvarez, Andre G Skirtach, Koen Raemdonck, Kevin Braeckmans
Journal
Small
Abstract
Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention f (show more...)Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention for the detection of cancer at an early stage. In this study the feasibility of using a surface enhanced Raman spectroscopy (SERS) based method to distinguish between ELVs derived from different cellular origins is evaluated. A gold nanoparticle based shell is deposited on the surface of ELVs derived from cancerous and healthy cells, which enhances the Raman signal while maintaining a colloidal suspension of individual vesicles. This nanocoating allows the recording of SERS spectra from single vesicles. By using partial least squares discriminant analysis on the obtained spectra, vesicles from different origin can be distinguished, even when present in the same mixture. This proof-of-concept study paves the way for noninvasive (cancer) diagnostic tools based on exosomal SERS fingerprinting in combination with multivariate statistical analysis. (hide)
EV-METRIC
78% (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
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
Cell Name
B16F10
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.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
DG
(d)(U)C
UF
Protein markers
EV: CD81/ HSP70/ CD63/ ?-actin
non-EV: None
Proteomics
no
EV density (g/ml)
1.14
Show all info
Study aim
Biomarker/Technical analysis comparing/optimizing EV-­related methods/New methodological development
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
B16F10
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Other preparation;Ultrafiltration
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
12.5%
Highest density fraction
50%
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 55 Ti
Speed (g)
150000
Duration (min)
900
Fraction volume (mL)
0.5
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
150000
Ultra filtration
Cut-off size (kDa)
30
Membrane type
Not specified
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63/ ?-actin/ HSP70/ CD81
Flow cytometry
Hardware adjustments
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
120
Particle yield
Not reported
EM
EM-type
Cryo-­EM
Image type
Wide-field
EV concentration
Not
1 - 2 of 2