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Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation

The aim of the study was to explore the polyethylene glycol–dextran two-phase polymer system formed in human plasma to isolate the exosome-enriched fraction of plasma extracellular nanovesicles (ENVs). Systematic analysis was performed to determine the optimal combination of the polymer mixture para...

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Autores principales: Slyusarenko, Maria, Nikiforova, Nadezhda, Sidina, Elena, Nazarova, Inga, Egorov, Vladimir, Garmay, Yuri, Merdalimova, Anastasiia, Yevlampieva, Natalia, Gorin, Dmitry, Malek, Anastasia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7867002/
https://www.ncbi.nlm.nih.gov/pubmed/33572666
http://dx.doi.org/10.3390/polym13030458
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author Slyusarenko, Maria
Nikiforova, Nadezhda
Sidina, Elena
Nazarova, Inga
Egorov, Vladimir
Garmay, Yuri
Merdalimova, Anastasiia
Yevlampieva, Natalia
Gorin, Dmitry
Malek, Anastasia
author_facet Slyusarenko, Maria
Nikiforova, Nadezhda
Sidina, Elena
Nazarova, Inga
Egorov, Vladimir
Garmay, Yuri
Merdalimova, Anastasiia
Yevlampieva, Natalia
Gorin, Dmitry
Malek, Anastasia
author_sort Slyusarenko, Maria
collection PubMed
description The aim of the study was to explore the polyethylene glycol–dextran two-phase polymer system formed in human plasma to isolate the exosome-enriched fraction of plasma extracellular nanovesicles (ENVs). Systematic analysis was performed to determine the optimal combination of the polymer mixture parameters (molecular mass and concentration) that resulted in phase separation. The separated phases were analyzed by nanoparticle tracking analysis and Raman spectroscopy. The isolated vesicles were characterized by atomic force microscopy and dot blotting. In conclusion, the protein and microRNA contents of the isolated ENVs were assayed by flow cytometry and by reverse transcription followed by quantitative polymerase chain reaction (RT-qPCR), respectively. The presented results revealed the applicability of a new method for plasma ENV isolation and further analysis with a diagnostic purpose.
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spelling pubmed-78670022021-02-07 Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation Slyusarenko, Maria Nikiforova, Nadezhda Sidina, Elena Nazarova, Inga Egorov, Vladimir Garmay, Yuri Merdalimova, Anastasiia Yevlampieva, Natalia Gorin, Dmitry Malek, Anastasia Polymers (Basel) Article The aim of the study was to explore the polyethylene glycol–dextran two-phase polymer system formed in human plasma to isolate the exosome-enriched fraction of plasma extracellular nanovesicles (ENVs). Systematic analysis was performed to determine the optimal combination of the polymer mixture parameters (molecular mass and concentration) that resulted in phase separation. The separated phases were analyzed by nanoparticle tracking analysis and Raman spectroscopy. The isolated vesicles were characterized by atomic force microscopy and dot blotting. In conclusion, the protein and microRNA contents of the isolated ENVs were assayed by flow cytometry and by reverse transcription followed by quantitative polymerase chain reaction (RT-qPCR), respectively. The presented results revealed the applicability of a new method for plasma ENV isolation and further analysis with a diagnostic purpose. MDPI 2021-01-31 /pmc/articles/PMC7867002/ /pubmed/33572666 http://dx.doi.org/10.3390/polym13030458 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Slyusarenko, Maria
Nikiforova, Nadezhda
Sidina, Elena
Nazarova, Inga
Egorov, Vladimir
Garmay, Yuri
Merdalimova, Anastasiia
Yevlampieva, Natalia
Gorin, Dmitry
Malek, Anastasia
Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title_full Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title_fullStr Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title_full_unstemmed Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title_short Formation and Evaluation of a Two-Phase Polymer System in Human Plasma as a Method for Extracellular Nanovesicle Isolation
title_sort formation and evaluation of a two-phase polymer system in human plasma as a method for extracellular nanovesicle isolation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7867002/
https://www.ncbi.nlm.nih.gov/pubmed/33572666
http://dx.doi.org/10.3390/polym13030458
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