Título:
Characterization of extracellular vesicles and synthetic nanoparticles with four orthogonal single-particle analysis platforms
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Autor:
Tosar Rovira, Juan Pablo
Arab, Tanina
Mallick, Emily R.
Huang, Yiyao
Dong, Liang
Liao, Zhaohao
Arab, Tanina
Mallick, Emily R.
Huang, Yiyao
Dong, Liang
Liao, Zhaohao
Tutor:
Tipo de documento:
Artículo
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Palabras clave:
Ectosomes
Exosomes
Extracellular vesicles
Microvesicles
Nanoflow cytometry
Nanoparticle tracking analysis
Resistive pulse sensing
Single particle interferometric reflectance imaging sensing
Exosomes
Extracellular vesicles
Microvesicles
Nanoflow cytometry
Nanoparticle tracking analysis
Resistive pulse sensing
Single particle interferometric reflectance imaging sensing
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Año de publicación:
2021
Contenido:
Resumen:
We compared four orthogonal technologies for sizing, counting, and phenotyping of extracellular vesicles (EVs) and synthetic particles. The platforms were: singleparticle 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 andNFCMreturned similar particle counts,whileNTAdetected 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, SPIRIS, 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 fluorescencemode was able to detect at least twomarkers 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.
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Editorial:
International Society for Extracellular Vesicles
EN:
Journal of Extracellular Vesicles, 2021, 10(6): e12079.
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Citación:
Tosar Rovira, J, Arab, T, Mallick, E [y otros autores]. "Characterization of extracellular vesicles and synthetic nanoparticles with four orthogonal single-particle analysis platforms". Journal of Extracellular Vesicles. [en línea] 2021, 10(6): e12079. 20 h. Doi: 10.1002/jev2.12079.
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2001-3078
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Licencia Creative Commons Atribución (CC - By 4.0)
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| Ficheros | Descripción | Tamaño | Formato | ||
|---|---|---|---|---|---|
| 101002jev212079.pdf | — | 4.53 MB | Adobe PDF |