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Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles
We present a resource‐efficient approach to fabricate and operate a micro‐nanofluidic device that uses cross‐flow filtration to isolate and capture liposarcoma derived extracellular vesicles (EVs). The isolated extracellular vesicles were captured using EV‐specific protein markers to obtain vesicle...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887429/ https://www.ncbi.nlm.nih.gov/pubmed/33643547 http://dx.doi.org/10.1002/jev2.12062 |
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author | Casadei, Lucia Choudhury, Adarsh Sarchet, Patricia Mohana Sundaram, Prashanth Lopez, Gonzalo Braggio, Danielle Balakirsky, Gita Pollock, Raphael Prakash, Shaurya |
author_facet | Casadei, Lucia Choudhury, Adarsh Sarchet, Patricia Mohana Sundaram, Prashanth Lopez, Gonzalo Braggio, Danielle Balakirsky, Gita Pollock, Raphael Prakash, Shaurya |
author_sort | Casadei, Lucia |
collection | PubMed |
description | We present a resource‐efficient approach to fabricate and operate a micro‐nanofluidic device that uses cross‐flow filtration to isolate and capture liposarcoma derived extracellular vesicles (EVs). The isolated extracellular vesicles were captured using EV‐specific protein markers to obtain vesicle enriched media, which was then eluted for further analysis. Therefore, the micro‐nanofluidic device integrates the unit operations of size‐based separation with CD63 antibody immunoaffinity‐based capture of extracellular vesicles in the same device to evaluate EV‐cargo content for liposarcoma. The eluted media collected showed ∼76% extracellular vesicle recovery from the liposarcoma cell conditioned media and ∼32% extracellular vesicle recovery from dedifferentiated liposarcoma patient serum when compared against state‐of‐art extracellular vesicle isolation and subsequent quantification by ultracentrifugation. The results reported here also show a five‐fold increase in amount of critical liposarcoma‐relevant extracellular vesicle cargo obtained in 30 min presenting a significant advance over existing state‐of‐art. |
format | Online Article Text |
id | pubmed-7887429 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-78874292021-02-26 Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles Casadei, Lucia Choudhury, Adarsh Sarchet, Patricia Mohana Sundaram, Prashanth Lopez, Gonzalo Braggio, Danielle Balakirsky, Gita Pollock, Raphael Prakash, Shaurya J Extracell Vesicles Research Articles We present a resource‐efficient approach to fabricate and operate a micro‐nanofluidic device that uses cross‐flow filtration to isolate and capture liposarcoma derived extracellular vesicles (EVs). The isolated extracellular vesicles were captured using EV‐specific protein markers to obtain vesicle enriched media, which was then eluted for further analysis. Therefore, the micro‐nanofluidic device integrates the unit operations of size‐based separation with CD63 antibody immunoaffinity‐based capture of extracellular vesicles in the same device to evaluate EV‐cargo content for liposarcoma. The eluted media collected showed ∼76% extracellular vesicle recovery from the liposarcoma cell conditioned media and ∼32% extracellular vesicle recovery from dedifferentiated liposarcoma patient serum when compared against state‐of‐art extracellular vesicle isolation and subsequent quantification by ultracentrifugation. The results reported here also show a five‐fold increase in amount of critical liposarcoma‐relevant extracellular vesicle cargo obtained in 30 min presenting a significant advance over existing state‐of‐art. John Wiley and Sons Inc. 2021-02-16 2021-02 /pmc/articles/PMC7887429/ /pubmed/33643547 http://dx.doi.org/10.1002/jev2.12062 Text en © 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Casadei, Lucia Choudhury, Adarsh Sarchet, Patricia Mohana Sundaram, Prashanth Lopez, Gonzalo Braggio, Danielle Balakirsky, Gita Pollock, Raphael Prakash, Shaurya Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title | Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title_full | Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title_fullStr | Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title_full_unstemmed | Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title_short | Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
title_sort | cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887429/ https://www.ncbi.nlm.nih.gov/pubmed/33643547 http://dx.doi.org/10.1002/jev2.12062 |
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