Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.

Recent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and valida...

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Main Authors: Anirban Ghosh, Michelle Davey, Ian C Chute, Steven G Griffiths, Scott Lewis, Simi Chacko, David Barnett, Nicolas Crapoulet, Sébastien Fournier, Andrew Joy, Michelle C Caissie, Amanda D Ferguson, Melissa Daigle, M Vicki Meli, Stephen M Lewis, Rodney J Ouellette
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4201556?pdf=render
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spelling doaj-d1c92e7b12da4f40ba31ceeaf4faf0382020-11-24T21:35:12ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01910e11044310.1371/journal.pone.0110443Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.Anirban GhoshMichelle DaveyIan C ChuteSteven G GriffithsScott LewisSimi ChackoDavid BarnettNicolas CrapouletSébastien FournierAndrew JoyMichelle C CaissieAmanda D FergusonMelissa DaigleM Vicki MeliStephen M LewisRodney J OuelletteRecent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and validated a novel class of peptides (which we named venceremin, or Vn) that exhibit nucleotide-independent specific affinity for canonical heat shock proteins. The Vn peptides were validated to specifically and efficiently capture HSP-containing extracellular vesicles from cell culture growth media, plasma, and urine by electron microscopy, atomic force microscopy, sequencing of nucleic acid cargo, proteomic profiling, immunoblotting, and nanoparticle tracking analysis. All of these analyses confirmed the material captured by the Vn peptides was comparable to those purified by the standard ultracentrifugation method. We show that the Vn peptides are a useful tool for the rapid isolation of extracellular vesicles using standard laboratory equipment. Moreover, the Vn peptides are adaptable to diverse platforms and therefore represent an excellent solution to the challenge of extracellular vesicle isolation for research and clinical applications.http://europepmc.org/articles/PMC4201556?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Anirban Ghosh
Michelle Davey
Ian C Chute
Steven G Griffiths
Scott Lewis
Simi Chacko
David Barnett
Nicolas Crapoulet
Sébastien Fournier
Andrew Joy
Michelle C Caissie
Amanda D Ferguson
Melissa Daigle
M Vicki Meli
Stephen M Lewis
Rodney J Ouellette
spellingShingle Anirban Ghosh
Michelle Davey
Ian C Chute
Steven G Griffiths
Scott Lewis
Simi Chacko
David Barnett
Nicolas Crapoulet
Sébastien Fournier
Andrew Joy
Michelle C Caissie
Amanda D Ferguson
Melissa Daigle
M Vicki Meli
Stephen M Lewis
Rodney J Ouellette
Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
PLoS ONE
author_facet Anirban Ghosh
Michelle Davey
Ian C Chute
Steven G Griffiths
Scott Lewis
Simi Chacko
David Barnett
Nicolas Crapoulet
Sébastien Fournier
Andrew Joy
Michelle C Caissie
Amanda D Ferguson
Melissa Daigle
M Vicki Meli
Stephen M Lewis
Rodney J Ouellette
author_sort Anirban Ghosh
title Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
title_short Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
title_full Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
title_fullStr Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
title_full_unstemmed Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
title_sort rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2014-01-01
description Recent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and validated a novel class of peptides (which we named venceremin, or Vn) that exhibit nucleotide-independent specific affinity for canonical heat shock proteins. The Vn peptides were validated to specifically and efficiently capture HSP-containing extracellular vesicles from cell culture growth media, plasma, and urine by electron microscopy, atomic force microscopy, sequencing of nucleic acid cargo, proteomic profiling, immunoblotting, and nanoparticle tracking analysis. All of these analyses confirmed the material captured by the Vn peptides was comparable to those purified by the standard ultracentrifugation method. We show that the Vn peptides are a useful tool for the rapid isolation of extracellular vesicles using standard laboratory equipment. Moreover, the Vn peptides are adaptable to diverse platforms and therefore represent an excellent solution to the challenge of extracellular vesicle isolation for research and clinical applications.
url http://europepmc.org/articles/PMC4201556?pdf=render
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