PMID- 33850608
OWN - NLM
STAT- MEDLINE
DCOM- 20220325
LR  - 20220415
IS  - 2001-3078 (Print)
IS  - 2001-3078 (Electronic)
IS  - 2001-3078 (Linking)
VI  - 10
IP  - 6
DP  - 2021 Apr
TI  - Characterization of extracellular vesicles and synthetic nanoparticles with four 
      orthogonal single-particle analysis platforms.
PG  - e12079
LID - 10.1002/jev2.12079 [doi]
LID - e12079
AB  - We compared four orthogonal technologies for sizing, counting, and phenotyping of 
      extracellular vesicles (EVs) and synthetic particles. The platforms were: 
      single-particle interferometric reflectance imaging sensing (SP-IRIS) with 
      fluorescence, nanoparticle tracking analysis (NTA) with fluorescence, 
      microfluidic resistive pulse sensing (MRPS), and nanoflow cytometry measurement 
      (NFCM). EVs from the human T lymphocyte line H9 (high CD81, low CD63) and the 
      promonocytic line U937 (low CD81, high CD63) were separated from culture 
      conditioned medium (CCM) by differential ultracentrifugation (dUC) or a 
      combination of ultrafiltration (UF) and size exclusion chromatography (SEC) and 
      characterized by transmission electron microscopy (TEM) and Western blot (WB). 
      Mixtures of synthetic particles (silica and polystyrene spheres) with known sizes 
      and/or concentrations were also tested. MRPS and NFCM returned similar particle 
      counts, while NTA detected counts approximately one order of magnitude lower for 
      EVs, but not for synthetic particles. SP-IRIS events could not be used to 
      estimate particle concentrations. For sizing, SP-IRIS, MRPS, and NFCM returned 
      similar size profiles, with smaller sizes predominating (per power law 
      distribution), but with sensitivity typically dropping off below diameters of 60 
      nm. NTA detected a population of particles with a mode diameter greater than 100 
      nm. Additionally, SP-IRIS, MRPS, and NFCM were able to identify at least three of 
      four distinct size populations in a mixture of silica or polystyrene 
      nanoparticles. Finally, for tetraspanin phenotyping, the SP-IRIS platform in 
      fluorescence mode was able to detect at least two markers on the same particle, 
      while NFCM detected either CD81 or CD63. Based on the results of this study, we 
      can draw conclusions about existing single-particle analysis capabilities that 
      may be useful for EV biomarker development and mechanistic studies.
CI  - (c) 2021 The Authors. Journal of Extracellular Vesicles published by Wiley 
      Periodicals, LLC on behalf of the International Society for Extracellular 
      Vesicles.
FAU - Arab, Tanina
AU  - Arab T
AUID- ORCID: 0000-0003-4485-1096
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Mallick, Emily R
AU  - Mallick ER
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Huang, Yiyao
AU  - Huang Y
AUID- ORCID: 0000-0003-1749-963X
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Dong, Liang
AU  - Dong L
AUID- ORCID: 0000-0001-7689-3237
AD  - Department of Urology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
FAU - Liao, Zhaohao
AU  - Liao Z
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Zhao, Zezhou
AU  - Zhao Z
AUID- ORCID: 0000-0003-4792-6073
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Gololobova, Olesia
AU  - Gololobova O
AUID- ORCID: 0000-0002-0392-268X
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Smith, Barbara
AU  - Smith B
AD  - Department of Cell Biology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
FAU - Haughey, Norman J
AU  - Haughey NJ
AUID- ORCID: 0000-0001-5194-4122
AD  - Department of Neurology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
FAU - Pienta, Kenneth J
AU  - Pienta KJ
AUID- ORCID: 0000-0002-4138-2186
AD  - Department of Urology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
FAU - Slusher, Barbara S
AU  - Slusher BS
AUID- ORCID: 0000-0001-9814-4157
AD  - Department of Neurology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
AD  - Johns Hopkins Drug Discovery Johns Hopkins University School of Medicine 
      Baltimore Maryland USA.
FAU - Tarwater, Patrick M
AU  - Tarwater PM
AUID- ORCID: 0000-0002-6791-1090
AD  - Department of Epidemiology Johns Hopkins University Bloomberg School of Public 
      Health Baltimore Maryland USA.
FAU - Tosar, Juan Pablo
AU  - Tosar JP
AUID- ORCID: 0000-0002-2021-2479
AD  - Faculty of Science Universidad de la Republica Montevideo Uruguay.
AD  - Functional Genomics Unit Institut Pasteur de Montevideo Montevideo Uruguay.
FAU - Zivkovic, Angela M
AU  - Zivkovic AM
AUID- ORCID: 0000-0002-2828-7862
AD  - Department of Nutrition University of California Davis Davis California USA.
FAU - Vreeland, Wyatt N
AU  - Vreeland WN
AUID- ORCID: 0000-0003-0524-8403
AD  - Bioprocess Measurements Group National Institute of Standards and Technology 
      Gaithersburg Maryland USA.
FAU - Paulaitis, Michael E
AU  - Paulaitis ME
AUID- ORCID: 0000-0002-9628-1091
AD  - Center for Nanomedicine at the Wilmer Eye Institute Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
FAU - Witwer, Kenneth W
AU  - Witwer KW
AUID- ORCID: 0000-0003-1664-4233
AD  - Department of Molecular and Comparative Pathobiology Johns Hopkins University 
      School of Medicine Baltimore Maryland USA.
AD  - Department of Neurology Johns Hopkins University School of Medicine Baltimore 
      Maryland USA.
AD  - The Richman Family Precision Medicine Center of Excellence in Alzheimer's Disease 
      Johns Hopkins University School of Medicine Johns Hopkins Medicine and Johns 
      Hopkins Bayview Medical Center Baltimore Maryland USA.
LA  - eng
GR  - R33 MH118164/MH/NIMH NIH HHS/United States
GR  - UG3 CA241694/CA/NCI NIH HHS/United States
GR  - R01 AI144997/AI/NIAID NIH HHS/United States
GR  - R01 DA040385/DA/NIDA NIH HHS/United States
GR  - R01 DA047807/DA/NIDA NIH HHS/United States
GR  - R21 MH118164/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20210406
PL  - United States
TA  - J Extracell Vesicles
JT  - Journal of extracellular vesicles
JID - 101610479
RN  - 0 (Biomarkers)
RN  - 0 (Polystyrenes)
MH  - Biomarkers/analysis
MH  - Cell Line
MH  - Chromatography, Gel/methods
MH  - Extracellular Vesicles/*chemistry/*metabolism/*physiology
MH  - Humans
MH  - Microfluidics/methods
MH  - Microscopy, Electron, Transmission/methods
MH  - Nanoparticles/chemistry
MH  - Particle Size
MH  - Polystyrenes/analysis
MH  - Single Molecule Imaging/methods
MH  - Ultracentrifugation/methods
MH  - Ultrafiltration
PMC - PMC8023330
OTO - NOTNLM
OT  - ectosomes
OT  - exosomes
OT  - extracellular vesicles
OT  - microvesicles
OT  - nanoflow cytometry
OT  - nanoparticle tracking analysis
OT  - resistive pulse sensing
OT  - single particle interferometric reflectance imaging sensing
EDAT- 2021/04/15 06:00
MHDA- 2021/04/15 06:01
PMCR- 2021/04/06
CRDT- 2021/04/14 06:33
PHST- 2020/08/05 00:00 [received]
PHST- 2021/03/04 00:00 [revised]
PHST- 2021/03/05 00:00 [accepted]
PHST- 2021/04/14 06:33 [entrez]
PHST- 2021/04/15 06:00 [pubmed]
PHST- 2021/04/15 06:01 [medline]
PHST- 2021/04/06 00:00 [pmc-release]
AID - JEV212079 [pii]
AID - 10.1002/jev2.12079 [doi]
PST - ppublish
SO  - J Extracell Vesicles. 2021 Apr;10(6):e12079. doi: 10.1002/jev2.12079. Epub 2021 
      Apr 6.