Search > Results

You searched for: EV200078 (EV-TRACK ID)

Showing 1 - 1 of 1

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
EV200078 1/1 Homo sapiens saliva SEC (non-commercial)
Filtration
UF
Ogawa, Yuko 2021 75%

Study summary

Full title
All authors
Yuko Ogawa, Yoshihiro Akimoto, Mamoru Ikemoto, Yoshikuni Goto, Anna Ishikawa, Sakura Ohta, Yumi Takase, Hayato Kawakami, Masafumi Tsujimoto, Ryohei Yanoshita
Journal
Biochemistry and Biophysics Reports
Abstract
Background: Extracellular vesicles (EVs) have been isolated from various sources, including primary (show more...)Background: Extracellular vesicles (EVs) have been isolated from various sources, including primary and cultured cell lines and body fluids. Previous studies, including those conducted in our laboratory, have reported the stability of EVs under various storage conditions. Methods: EVs from human whole saliva were separated via size-exclusion chromatography. To simulate the effects of gastric or intestinal fluids on the stability of EVs, pepsin or pancreatin was added to the samples. Additionally, to determine the effect of bile acids, sodium cholate was added. The samples were then subjected to western blotting, dynamic light scattering, and transmission electron microscopy analyses. In addition, the activity of dipeptidyl peptidase (DPP) IV retained in the samples was examined to monitor the stability of EVs. Results: Under acidic conditions, with pepsin mimicking the milieu of the stomach, the EVs remained stable. However, they partially lost their membrane integrity in the presence of pancreatin and sodium cholate, indicating that they may be destabilized after passing through the duodenum. Although several associated proteins, such as mucin 5B and CD9 were degraded, DPP IV was stable, and its activity was retained under the simulated gastrointestinal conditions. Conclusion: Our data indicate that although EVs can pass through the stomach without undergoing significant damage, they may be disrupted in the intestine to release their contents. The consistent delivery of active components such as DPP IV from EVs into the intestine might play a role in the efficient modulation of homeostasis of the signal transduction pathways occurring in the gastrointestinal tract. (hide)
EV-METRIC
75% (98th 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
saliva
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
Size-exclusion chromatography (non-commercial)
Filtration
Ultrafiltration
Protein markers
EV: TSG101/ mucin 5B/ CD63/ CD81/ HSP90/ Alix/ Flotillin1/ IgA/ DPP IV/ HSP70/ CD9
non-EV: apolipoprotein B-100/ Albumin/ apolipoprotein A-1
Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
saliva
Sample Condition
Control condition
Separation Method
Filtration steps
> 0.45 µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Cellulose acetate
Size-exclusion chromatography
Total column volume (mL)
88
Sample volume/column (mL)
2
Resin type
Sephacryl S-500
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD9/ CD63/ DPP IV/ mucin 5B/ IgA/ HSP90/ TSG101/ HSP70/ Alix/ CD81
Not detected EV-associated proteins
Flotillin1
Detected contaminants
apolipoprotein A-1/ apolipoprotein B-100/ Albumin
Proteomics
Proteomics database
Yes:
Characterization: Particle analysis
DLS
Report type
Mean
Reported size (nm)
40
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
Report size (nm)
40
1 - 1 of 1
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV200078
species
Homo sapiens
sample type
saliva
condition
Control condition
separation protocol
Size-exclusion chromatography (non-commercial)
Filtration
Ultrafiltration
Exp. nr.
1
EV-METRIC %
75