Search > Results

You searched for: EV210210 (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
EV210210 1/1 Homo sapiens Expi293F (d)(U)C
Tangential flow filtration
qEV10
Filtration
UF
Driedonks T 2022 63%

Study summary

Full title
All authors
Driedonks T, Jiang L, Carlson B, Han Z, Liu G, Queen SE, Shirk EN, Gololobova O, Liao Z, Nyberg LH, Lima G, Paniushkina L, Garcia-Contreras M, Schonvisky K, Castell N, Stover M, Guerrero-Martin S, Richardson R, Smith B, Machairaki V, Lai CP, Izzi JM, Hutchinson EK, Pate KAM, Witwer KW
Journal
J Extracell Biol
Abstract
Extracellular vesicles (EVs) have potential in disease treatment since they can be loaded with thera (show more...)Extracellular vesicles (EVs) have potential in disease treatment since they can be loaded with therapeutic molecules and engineered for retention by specific tissues. However, questions remain on optimal dosing, administration, and pharmacokinetics. Previous studies have addressed biodistribution and pharmacokinetics in rodents, but little evidence is available for larger animals. Here, we investigated the pharmacokinetics and biodistribution of Expi293F-derived EVs labelled with a highly sensitive nanoluciferase reporter (palmGRET) in a non-human primate model (Macaca nemestrina), comparing intravenous (IV) and intranasal (IN) administration over a 125-fold dose range. We report that EVs administered IV had longer circulation times in plasma than previously reported in mice and were detectable in cerebrospinal fluid (CSF) after 30-60 minutes. EV association with PBMCs, especially B-cells, was observed as early as one minute post-administration. EVs were detected in liver and spleen within one hour of IV administration. However, IN delivery was minimal, suggesting that pretreatment approaches may be needed in large animals. Furthermore, EV circulation times strongly decreased after repeated IV administration, possibly due to immune responses and with clear implications for xenogeneic EV-based therapeutics. We hope that our findings from this baseline study in macaques will help to inform future research and therapeutic development of EVs. (hide)
EV-METRIC
63% (92nd 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
pLenti-palmGRET (Addgene #158221) transient expression
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
Tangential flow filtration
Commercial method
Filtration
Ultrafiltration
Protein markers
EV: CD9/ CD63/ CD81/ TSG101/ CD29/ CD146/ CD326/ CD44/ ROR1/ CD24
non-EV: Calnexin
Proteomics
no
Show all info
Study aim
Biodistribution/pharmacokinetics/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
Expi293F
Cell count
3.30E+09
Separation Method
Filtration steps
0.2 or 0.22 ┬Ám
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Regenerated cellulose
Commercial kit
qEV10
Other
Name other separation method
Tangential flow filtration
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Antibody details provided?
No
Detected EV-associated proteins
CD9/ CD63/ TSG101/ CD47
Not detected contaminants
Calnexin
Other 1
SP-IRIS (Exoview)
Detected EV-associated proteins
CD63/ CD81
Other 2
MACSPlex
Detected EV-associated proteins
CD9/ CD63/ CD81/ CD29/ CD146/ CD326/ CD44/ ROR1/ CD24
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
122.6
EV concentration
Yes
Particle analysis: flow cytometry
Flow cytometer type
Amnis ImageStream MKII
Hardware adjustment
EM
EM-type
Transmission-EM
Image type
Wide-field
Extra information
Size-exclusion chromatography details: Izon qEV10, first 20 ml after the void fraction were considered EV-enriched fractions/ Tangential Flow Filtration: Sartorius Vivaflow 50R cassettes, 100 kD
1 - 1 of 1
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV210210
species
Homo sapiens
sample type
Cell culture
cell type
Expi293F
condition
pLenti-palmGRET
(Addgene #158221)
transient expression
separation protocol
dUC/ Tangential flow
filtration/ qEV10/ Filtration/
Ultrafiltration
Exp. nr.
1
EV-METRIC %
63