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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...

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Autores principales: Casadei, Lucia, Choudhury, Adarsh, Sarchet, Patricia, Mohana Sundaram, Prashanth, Lopez, Gonzalo, Braggio, Danielle, Balakirsky, Gita, Pollock, Raphael, Prakash, Shaurya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
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.
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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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