Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles
Abstract 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...
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Series: | Journal of Extracellular Vesicles |
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Online Access: | https://doi.org/10.1002/jev2.12062 |
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doaj-ede2998c3b5143528ae6ecc374830a1a2021-04-20T13:45:19ZengTaylor & Francis GroupJournal of Extracellular Vesicles2001-30782021-02-01104n/an/a10.1002/jev2.12062Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesiclesLucia Casadei0Adarsh Choudhury1Patricia Sarchet2Prashanth Mohana Sundaram3Gonzalo Lopez4Danielle Braggio5Gita Balakirsky6Raphael Pollock7Shaurya Prakash8Comprehensive Cancer Center The Ohio State University Columbus Ohio USADepartment of Mechanical and Aerospace Engineering The Ohio State University Columbus Ohio USAComprehensive Cancer Center The Ohio State University Columbus Ohio USADepartment of Mechanical and Aerospace Engineering The Ohio State University Columbus Ohio USAComprehensive Cancer Center The Ohio State University Columbus Ohio USAComprehensive Cancer Center The Ohio State University Columbus Ohio USAComprehensive Cancer Center The Ohio State University Columbus Ohio USADepartment of Mechanical and Aerospace Engineering The Ohio State University Columbus Ohio USAComprehensive Cancer Center The Ohio State University Columbus Ohio USAAbstract 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.https://doi.org/10.1002/jev2.12062cross flow filtrationDNAeEVextracellular vesicleslEVliposarcoma |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lucia Casadei Adarsh Choudhury Patricia Sarchet Prashanth Mohana Sundaram Gonzalo Lopez Danielle Braggio Gita Balakirsky Raphael Pollock Shaurya Prakash |
spellingShingle |
Lucia Casadei Adarsh Choudhury Patricia Sarchet Prashanth Mohana Sundaram Gonzalo Lopez Danielle Braggio Gita Balakirsky Raphael Pollock Shaurya Prakash Cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles Journal of Extracellular Vesicles cross flow filtration DNA eEV extracellular vesicles lEV liposarcoma |
author_facet |
Lucia Casadei Adarsh Choudhury Patricia Sarchet Prashanth Mohana Sundaram Gonzalo Lopez Danielle Braggio Gita Balakirsky Raphael Pollock Shaurya Prakash |
author_sort |
Lucia Casadei |
title |
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_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_sort |
cross‐flow microfiltration for isolation, selective capture and release of liposarcoma extracellular vesicles |
publisher |
Taylor & Francis Group |
series |
Journal of Extracellular Vesicles |
issn |
2001-3078 |
publishDate |
2021-02-01 |
description |
Abstract 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. |
topic |
cross flow filtration DNA eEV extracellular vesicles lEV liposarcoma |
url |
https://doi.org/10.1002/jev2.12062 |
work_keys_str_mv |
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