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Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids

Extracellular vesicles (EVs), such as exosomes, are nanovesicles that have received significant attention due to their ability to contain various molecular cargos. EVs found in biological fluids have been demonstrated to have therapeutic potential, including as biomarkers. Despite being extensively...

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Autores principales: Chandrasekera, Dhananjie, Shah, Rishi, van Hout, Isabelle, De Jonge, Willow, Bunton, Richard, Parry, Dominic, Davis, Philip, Katare, Rajesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Ivyspring International Publisher 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10161387/
https://www.ncbi.nlm.nih.gov/pubmed/37151803
http://dx.doi.org/10.7150/ntno.82939
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author Chandrasekera, Dhananjie
Shah, Rishi
van Hout, Isabelle
De Jonge, Willow
Bunton, Richard
Parry, Dominic
Davis, Philip
Katare, Rajesh
author_facet Chandrasekera, Dhananjie
Shah, Rishi
van Hout, Isabelle
De Jonge, Willow
Bunton, Richard
Parry, Dominic
Davis, Philip
Katare, Rajesh
author_sort Chandrasekera, Dhananjie
collection PubMed
description Extracellular vesicles (EVs), such as exosomes, are nanovesicles that have received significant attention due to their ability to contain various molecular cargos. EVs found in biological fluids have been demonstrated to have therapeutic potential, including as biomarkers. Despite being extensively studied, a significant downfall in EV research is the lack of standardised protocol for its isolation from human biological fluids, where EVs usually exist at low densities. In this study, we tested two well-established EV isolation protocols, precipitation, and size exclusion chromatography (SEC), to determine their efficiency in isolating EVs from the pericardial fluid. Precipitation alone resulted in high yields of low-purity exosomes as tested by DLS analysis, transmission electron microscopy, immunogold labelling and western blotting for the exosomal surface proteins. While EVs isolated by SEC were pure, the concentration was low. Interestingly, the combination of precipitation followed by SEC resulted in high EV yields with good purity. Our results suggest that the combination method can be adapted to isolate EVs from body fluids which have low densities of EV.
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spelling pubmed-101613872023-05-06 Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids Chandrasekera, Dhananjie Shah, Rishi van Hout, Isabelle De Jonge, Willow Bunton, Richard Parry, Dominic Davis, Philip Katare, Rajesh Nanotheranostics Research Paper Extracellular vesicles (EVs), such as exosomes, are nanovesicles that have received significant attention due to their ability to contain various molecular cargos. EVs found in biological fluids have been demonstrated to have therapeutic potential, including as biomarkers. Despite being extensively studied, a significant downfall in EV research is the lack of standardised protocol for its isolation from human biological fluids, where EVs usually exist at low densities. In this study, we tested two well-established EV isolation protocols, precipitation, and size exclusion chromatography (SEC), to determine their efficiency in isolating EVs from the pericardial fluid. Precipitation alone resulted in high yields of low-purity exosomes as tested by DLS analysis, transmission electron microscopy, immunogold labelling and western blotting for the exosomal surface proteins. While EVs isolated by SEC were pure, the concentration was low. Interestingly, the combination of precipitation followed by SEC resulted in high EV yields with good purity. Our results suggest that the combination method can be adapted to isolate EVs from body fluids which have low densities of EV. Ivyspring International Publisher 2023-04-02 /pmc/articles/PMC10161387/ /pubmed/37151803 http://dx.doi.org/10.7150/ntno.82939 Text en © The author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.
spellingShingle Research Paper
Chandrasekera, Dhananjie
Shah, Rishi
van Hout, Isabelle
De Jonge, Willow
Bunton, Richard
Parry, Dominic
Davis, Philip
Katare, Rajesh
Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title_full Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title_fullStr Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title_full_unstemmed Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title_short Combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
title_sort combination of precipitation and size exclusion chromatography as an effective method for exosome like extracellular vesicle isolation from pericardial fluids
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10161387/
https://www.ncbi.nlm.nih.gov/pubmed/37151803
http://dx.doi.org/10.7150/ntno.82939
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