Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry

Acetone precipitation was evaluated as a rapid, simple, low-cost, and efficient method for the selective purification of O-glycopeptides from enzymatic digests of glycoproteins. Ovalbumin (OVA), human and bovine α1-acid glycoprotein (hAGP and bAGP), human apolipoprotein C–III (APO-C3), and recombina...

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Detalhes bibliográficos
Autores: Mancera Arteu, Montserrat, Giménez López, Estela, Benavente Moreno, Fernando J. (Julián), Barbosa Torralbo, José, Sanz Nebot, María Victoria
Tipo de documento: artigo
Estado:Versión aceptada para publicación
Data de publicação:2017
País:España
Recursos:Universidad de Barcelona
Repositório:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/225657
Acesso em linha:https://hdl.handle.net/2445/225657
Access Level:Acceso aberto
Palavra-chave:Acetona
Cetones
Electroforesi capil·lar
Acetone
Ketones
Capillary electrophoresis
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spelling Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometryMancera Arteu, MontserratGiménez López, EstelaBenavente Moreno, Fernando J. (Julián)Barbosa Torralbo, JoséSanz Nebot, María VictoriaAcetonaCetonesElectroforesi capil·larAcetoneKetonesCapillary electrophoresisAcetone precipitation was evaluated as a rapid, simple, low-cost, and efficient method for the selective purification of O-glycopeptides from enzymatic digests of glycoproteins. Ovalbumin (OVA), human and bovine α1-acid glycoprotein (hAGP and bAGP), human apolipoprotein C–III (APO-C3), and recombinant human erythropoietin (rhEPO) were used to obtain enzymatic digests with a broad and varied set of peptides, N-glycopeptides, and O-glycopeptides. After digestion and before capillary electrophoresis mass spectrometry (CE-MS) analysis, the amount of ice-cold acetone added to the digests was optimized to maximize recoveries of O-glycopeptides. Furthermore, the different behavior of peptides, N- and O-glycopeptides was explained by studying with multivariate data analysis methods the influence of several physicochemical parameters and properties related to their composition and structure. Principal component analysis (PCA) and, afterward, partial least-squares discriminant analysis (PLS-DA) were used to identify the most significant variables and their importance to differentiate between peptides, N-glycopeptides and O-glycopeptides, or within these classes. This information was useful to understand precipitation of these compounds after addition of acetone and for the selection of the optimal conditions for purification of specific O-glycopeptide biomarkers. Special attention was paid to O126-glycopeptide glycoforms of rhEPO because of their applicability in biopharmaceutical quality control and doping analysis.American Chemical Society2017info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttps://hdl.handle.net/2445/225657Articles publicats en revistes (Enginyeria Química i Química Analítica)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésVersió postprint del document publicat a: https://doi.org/10.1021/acs.jproteome.7b00524Journal of Proteome Research, 2017, vol. 16, p. 4166-4176https://doi.org/10.1021/acs.jproteome.7b00524(c) American Chemical Society, 2017info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2256572026-05-27T06:46:51Z
dc.title.none.fl_str_mv Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
title Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
spellingShingle Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
Mancera Arteu, Montserrat
Acetona
Cetones
Electroforesi capil·lar
Acetone
Ketones
Capillary electrophoresis
title_short Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
title_full Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
title_fullStr Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
title_full_unstemmed Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
title_sort Analysis of O-glycopeptides by acetone enrichment and capillary electrophoresis-mass spectrometry
dc.creator.none.fl_str_mv Mancera Arteu, Montserrat
Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
Barbosa Torralbo, José
Sanz Nebot, María Victoria
author Mancera Arteu, Montserrat
author_facet Mancera Arteu, Montserrat
Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
Barbosa Torralbo, José
Sanz Nebot, María Victoria
author_role author
author2 Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
Barbosa Torralbo, José
Sanz Nebot, María Victoria
author2_role author
author
author
author
dc.subject.none.fl_str_mv Acetona
Cetones
Electroforesi capil·lar
Acetone
Ketones
Capillary electrophoresis
topic Acetona
Cetones
Electroforesi capil·lar
Acetone
Ketones
Capillary electrophoresis
description Acetone precipitation was evaluated as a rapid, simple, low-cost, and efficient method for the selective purification of O-glycopeptides from enzymatic digests of glycoproteins. Ovalbumin (OVA), human and bovine α1-acid glycoprotein (hAGP and bAGP), human apolipoprotein C–III (APO-C3), and recombinant human erythropoietin (rhEPO) were used to obtain enzymatic digests with a broad and varied set of peptides, N-glycopeptides, and O-glycopeptides. After digestion and before capillary electrophoresis mass spectrometry (CE-MS) analysis, the amount of ice-cold acetone added to the digests was optimized to maximize recoveries of O-glycopeptides. Furthermore, the different behavior of peptides, N- and O-glycopeptides was explained by studying with multivariate data analysis methods the influence of several physicochemical parameters and properties related to their composition and structure. Principal component analysis (PCA) and, afterward, partial least-squares discriminant analysis (PLS-DA) were used to identify the most significant variables and their importance to differentiate between peptides, N-glycopeptides and O-glycopeptides, or within these classes. This information was useful to understand precipitation of these compounds after addition of acetone and for the selection of the optimal conditions for purification of specific O-glycopeptide biomarkers. Special attention was paid to O126-glycopeptide glycoforms of rhEPO because of their applicability in biopharmaceutical quality control and doping analysis.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/225657
url https://hdl.handle.net/2445/225657
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Versió postprint del document publicat a: https://doi.org/10.1021/acs.jproteome.7b00524
Journal of Proteome Research, 2017, vol. 16, p. 4166-4176
https://doi.org/10.1021/acs.jproteome.7b00524
dc.rights.none.fl_str_mv (c) American Chemical Society, 2017
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Chemical Society, 2017
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv Articles publicats en revistes (Enginyeria Química i Química Analítica)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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