Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry

The aim of this study is to present a comparative study of different methodologies for the extraction of proteins from platelet-derived extracellular vesicles (pEVs) prior to subsequent mass spectrometry (MS) analysis. pEVs were isolated by size exclusion chromatography (SEC) from human platelet lys...

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Autores: Ráez-Meseguer, Carmen, Amengual-Tugores, Andreu Miquel, Forteza-Genestra, Maria Antònia, Orvay-Pintos, Francisca, Gomila, Rosa M, Martorell-Crespí, Gabriel, Calvo Benito, Javier, Gayà, Antoni, Monjo, Marta, Ramis, Joana Maria
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Conselleria de Salut i Consum del Govern de les Illes Balears
Repositorio:Docusalut
Idioma:inglés
OAI Identifier:oai:docusalut.com:20.500.13003/25472
Acceso en línea:https://hdl.handle.net/20.500.13003/25472
Access Level:acceso abierto
Palabra clave:Extracellular Vesicles
Mass Spectrometry
Proteomics
Vesículas Extracelulares
Espectrometría de Masas
Proteómica
extracellular vesicles
mass spectrometry
methodologies
platelets
proteins
proteomics
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spelling Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass SpectrometryRáez-Meseguer, CarmenAmengual-Tugores, Andreu MiquelForteza-Genestra, Maria AntòniaOrvay-Pintos, FranciscaGomila, Rosa MMartorell-Crespí, GabrielCalvo Benito, JavierGayà, AntoniMonjo, MartaRamis, Joana MariaExtracellular VesiclesMass SpectrometryProteomicsVesículas ExtracelularesEspectrometría de MasasProteómicaextracellular vesiclesmass spectrometrymethodologiesplateletsproteinsproteomicsThe aim of this study is to present a comparative study of different methodologies for the extraction of proteins from platelet-derived extracellular vesicles (pEVs) prior to subsequent mass spectrometry (MS) analysis. pEVs were isolated by size exclusion chromatography (SEC) from human platelet lysates (PL) and characterized by identifying specific markers by Western blot, visualizing morphology by transmission electron microscopy (TEM) and analyzing concentration and size via nanoparticle tracking analysis (NTA). Protein isolation was performed through three different methodologies based on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), organic solvent precipitation (OSP), or magnetic beads (MB), followed by protein digestion and sample acquisition by LC-MS/MS. Clustering of the samples according to methodology is observed in the principal component analysis (PCA), although no significant differences in terms of normalized abundances are reached. Similarly, a small number of proteins were identified as unique by each methodology, with 91.3% coincidence among all three procedures. In addition, the bioinformatic results of the enrichment analysis and the numbers of proteins already identified in the Vesiclepedia database are highly similar for the three methodologies. Overall, all three methodologies analyzed are optimal for the extraction of proteins from pEV and could be considered according to their intrinsic characteristics, in accordance with the research requirements.20252025-07-2120252025-07-21research articlehttp://purl.org/coar/resource_type/c_2df8fbb1info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.13003/25472reponame:Docusalutinstname:Conselleria de Salut i Consum del Govern de les Illes BalearsInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:docusalut.com:20.500.13003/254722026-06-22T12:44:07Z
dc.title.none.fl_str_mv Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
title Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
spellingShingle Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
Ráez-Meseguer, Carmen
Extracellular Vesicles
Mass Spectrometry
Proteomics
Vesículas Extracelulares
Espectrometría de Masas
Proteómica
extracellular vesicles
mass spectrometry
methodologies
platelets
proteins
proteomics
title_short Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
title_full Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
title_fullStr Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
title_full_unstemmed Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
title_sort Comparative Analysis of Platelet-Derived Extracellular Vesicle Protein Extraction Methodologies for Mass Spectrometry
dc.creator.none.fl_str_mv Ráez-Meseguer, Carmen
Amengual-Tugores, Andreu Miquel
Forteza-Genestra, Maria Antònia
Orvay-Pintos, Francisca
Gomila, Rosa M
Martorell-Crespí, Gabriel
Calvo Benito, Javier
Gayà, Antoni
Monjo, Marta
Ramis, Joana Maria
author Ráez-Meseguer, Carmen
author_facet Ráez-Meseguer, Carmen
Amengual-Tugores, Andreu Miquel
Forteza-Genestra, Maria Antònia
Orvay-Pintos, Francisca
Gomila, Rosa M
Martorell-Crespí, Gabriel
Calvo Benito, Javier
Gayà, Antoni
Monjo, Marta
Ramis, Joana Maria
author_role author
author2 Amengual-Tugores, Andreu Miquel
Forteza-Genestra, Maria Antònia
Orvay-Pintos, Francisca
Gomila, Rosa M
Martorell-Crespí, Gabriel
Calvo Benito, Javier
Gayà, Antoni
Monjo, Marta
Ramis, Joana Maria
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv
dc.subject.none.fl_str_mv Extracellular Vesicles
Mass Spectrometry
Proteomics
Vesículas Extracelulares
Espectrometría de Masas
Proteómica
extracellular vesicles
mass spectrometry
methodologies
platelets
proteins
proteomics
topic Extracellular Vesicles
Mass Spectrometry
Proteomics
Vesículas Extracelulares
Espectrometría de Masas
Proteómica
extracellular vesicles
mass spectrometry
methodologies
platelets
proteins
proteomics
description The aim of this study is to present a comparative study of different methodologies for the extraction of proteins from platelet-derived extracellular vesicles (pEVs) prior to subsequent mass spectrometry (MS) analysis. pEVs were isolated by size exclusion chromatography (SEC) from human platelet lysates (PL) and characterized by identifying specific markers by Western blot, visualizing morphology by transmission electron microscopy (TEM) and analyzing concentration and size via nanoparticle tracking analysis (NTA). Protein isolation was performed through three different methodologies based on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), organic solvent precipitation (OSP), or magnetic beads (MB), followed by protein digestion and sample acquisition by LC-MS/MS. Clustering of the samples according to methodology is observed in the principal component analysis (PCA), although no significant differences in terms of normalized abundances are reached. Similarly, a small number of proteins were identified as unique by each methodology, with 91.3% coincidence among all three procedures. In addition, the bioinformatic results of the enrichment analysis and the numbers of proteins already identified in the Vesiclepedia database are highly similar for the three methodologies. Overall, all three methodologies analyzed are optimal for the extraction of proteins from pEV and could be considered according to their intrinsic characteristics, in accordance with the research requirements.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025-07-21
2025
2025-07-21
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.13003/25472
url https://hdl.handle.net/20.500.13003/25472
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Docusalut
instname:Conselleria de Salut i Consum del Govern de les Illes Balears
instname_str Conselleria de Salut i Consum del Govern de les Illes Balears
reponame_str Docusalut
collection Docusalut
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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