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Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application

Background: Exosomes are ubiquitous extracellular nanovesicles secreted from almost all living cells that are thought to be involved in several important cellular processes, including cell–cell communication and signaling. Exosomes serve as a liquid biopsy tool for clinical and translational researc...

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Autores principales: Tangwattanachuleeporn, Marut, Muanwien, Phijitra, Teethaisong, Yothin, Somparn, Poorichya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9694916/
https://www.ncbi.nlm.nih.gov/pubmed/36363557
http://dx.doi.org/10.3390/medicina58111600
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author Tangwattanachuleeporn, Marut
Muanwien, Phijitra
Teethaisong, Yothin
Somparn, Poorichya
author_facet Tangwattanachuleeporn, Marut
Muanwien, Phijitra
Teethaisong, Yothin
Somparn, Poorichya
author_sort Tangwattanachuleeporn, Marut
collection PubMed
description Background: Exosomes are ubiquitous extracellular nanovesicles secreted from almost all living cells that are thought to be involved in several important cellular processes, including cell–cell communication and signaling. Exosomes serve as a liquid biopsy tool for clinical and translational research. Although many techniques have been used to isolate exosomes, including ultracentrigation, size-exclusion chromatography, and immunocapturing-based techniques, these techniques are not convenient, they require expensive instrumentation, and they are unhandy for clinical samples. Precipitation techniques from available commercial kits that contain polyethelene glycol (PEG) are now widely used, but these kits are expensive, especially if a large number of biological samples are to be processed. Objective: the purpose of this study is to compare and optimize the efficacy of different concentrations of PEG with two commercial kits ExoQuick (SBI) and Total Exosome Isolation (TEI) from Invitrogen in human plasma. Methods and Materials: we determined exosome quantity, size distribution, marker expression, and downstream application. Results: among the precipitation methods, we found the size of particles and concentrations with 10–20% PEG are similar to ExoQuick and better than TEI. Interestingly, we detected cfDNA with ExoQuick and 10–20% PEG but not TEI and 5% PEG. Moreover, 10% PEG detection of miR-122 and miR-16 expression was superior to ExoQuick and TEI. Furthermore, in proteomics results it also found the identified proteins better than commercial kits but there was a high level of contamination of other proteins in serum. Conclusions: together, these findings show that an optimal concentration of 10% PEG serves as a guide for use with clinical samples in exosome isolation for downstream applications.
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spelling pubmed-96949162022-11-26 Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application Tangwattanachuleeporn, Marut Muanwien, Phijitra Teethaisong, Yothin Somparn, Poorichya Medicina (Kaunas) Article Background: Exosomes are ubiquitous extracellular nanovesicles secreted from almost all living cells that are thought to be involved in several important cellular processes, including cell–cell communication and signaling. Exosomes serve as a liquid biopsy tool for clinical and translational research. Although many techniques have been used to isolate exosomes, including ultracentrigation, size-exclusion chromatography, and immunocapturing-based techniques, these techniques are not convenient, they require expensive instrumentation, and they are unhandy for clinical samples. Precipitation techniques from available commercial kits that contain polyethelene glycol (PEG) are now widely used, but these kits are expensive, especially if a large number of biological samples are to be processed. Objective: the purpose of this study is to compare and optimize the efficacy of different concentrations of PEG with two commercial kits ExoQuick (SBI) and Total Exosome Isolation (TEI) from Invitrogen in human plasma. Methods and Materials: we determined exosome quantity, size distribution, marker expression, and downstream application. Results: among the precipitation methods, we found the size of particles and concentrations with 10–20% PEG are similar to ExoQuick and better than TEI. Interestingly, we detected cfDNA with ExoQuick and 10–20% PEG but not TEI and 5% PEG. Moreover, 10% PEG detection of miR-122 and miR-16 expression was superior to ExoQuick and TEI. Furthermore, in proteomics results it also found the identified proteins better than commercial kits but there was a high level of contamination of other proteins in serum. Conclusions: together, these findings show that an optimal concentration of 10% PEG serves as a guide for use with clinical samples in exosome isolation for downstream applications. MDPI 2022-11-04 /pmc/articles/PMC9694916/ /pubmed/36363557 http://dx.doi.org/10.3390/medicina58111600 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tangwattanachuleeporn, Marut
Muanwien, Phijitra
Teethaisong, Yothin
Somparn, Poorichya
Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title_full Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title_fullStr Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title_full_unstemmed Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title_short Optimizing Concentration of Polyethelene Glycol for Exosome Isolation from Plasma for Downstream Application
title_sort optimizing concentration of polyethelene glycol for exosome isolation from plasma for downstream application
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9694916/
https://www.ncbi.nlm.nih.gov/pubmed/36363557
http://dx.doi.org/10.3390/medicina58111600
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