Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry

In this study, several chromatographic sorbents: porous graphitic carbon (PGC), aminopropyl hydrophilic interaction (aminopropyl-HILIC), and phenylboronic acid (PBA) were assessed for the analysis of glycopeptides by on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CEM...

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Autores: Mancera Arteu, Montserrat, Benavente Moreno, Fernando J. (Julián), Sanz Nebot, María Victoria, Giménez López, Estela
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/199374
Acceso en línea:https://hdl.handle.net/2445/199374
Access Level:acceso abierto
Palabra clave:Electroforesi capil·lar
Espectrometria de masses
Glicopèptids
Capillary electrophoresis
Mass spectrometry
Glycopeptides
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spelling Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometryMancera Arteu, MontserratBenavente Moreno, Fernando J. (Julián)Sanz Nebot, María VictoriaGiménez López, EstelaElectroforesi capil·larEspectrometria de massesGlicopèptidsCapillary electrophoresisMass spectrometryGlycopeptidesIn this study, several chromatographic sorbents: porous graphitic carbon (PGC), aminopropyl hydrophilic interaction (aminopropyl-HILIC), and phenylboronic acid (PBA) were assessed for the analysis of glycopeptides by on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CEMS). As the PBA sorbent provided the most promising results, a PBA-SPE-CE-MS method was developed for the selective and sensitive preconcentration of glycopeptides from enzymatic digests of glycoproteins. Recombinant human erythropoietin (rhEPO) was selected as the model glycoprotein and subjected to enzymatic digestion with several proteases. The tryptic O126 and N83 glycopeptides from rhEPO were targeted to optimize the methodology. Under the optimized conditions, intraday precision, linearity, limits of detection (LODs), and microcartridge lifetime were evaluated, obtaining improved results compared to that from a previously reported TiO2-SPE-CE-MS method, especially for LODs of N-glycopeptides (up to 500 times lower than by CE-MS and up to 200 times lower than by TiO2-SPE-CE-MS). Moreover, rhEPO Glu-C digests were also analyzed by PBA-SPE-CE-MS to better characterize N24 and N38 glycopeptides. Finally, the established method was used to analyze two rhEPO products (EPOCIM and NeuroEPO plus), demonstrating its applicability in biopharmaceutical analysis. The sensitivity of the proposed PBA-SPE-CEMS method improves the existing CE-MS methodologies for glycopeptide analysis and shows a great potential in glycoprotein analysis to deeply characterize protein glycosites even at low concentrations of the protein digest.American Chemical Society2022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/199374Articles publicats en revistes (Enginyeria Química i Química Analítica)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1021/acs.jproteome.2c00569Journal of Proteome Research, 2022, vol. 22, p. 826-836https://doi.org/10.1021/acs.jproteome.2c00569cc-by (c) Mancera Arteu, Montserrat et al. , 2022http://creativecommons.org/licenses/by/3.0/es/info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1993742026-05-27T06:46:51Z
dc.title.none.fl_str_mv Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
title Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
spellingShingle Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
Mancera Arteu, Montserrat
Electroforesi capil·lar
Espectrometria de masses
Glicopèptids
Capillary electrophoresis
Mass spectrometry
Glycopeptides
title_short Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
title_full Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
title_fullStr Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
title_full_unstemmed Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
title_sort Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry
dc.creator.none.fl_str_mv Mancera Arteu, Montserrat
Benavente Moreno, Fernando J. (Julián)
Sanz Nebot, María Victoria
Giménez López, Estela
author Mancera Arteu, Montserrat
author_facet Mancera Arteu, Montserrat
Benavente Moreno, Fernando J. (Julián)
Sanz Nebot, María Victoria
Giménez López, Estela
author_role author
author2 Benavente Moreno, Fernando J. (Julián)
Sanz Nebot, María Victoria
Giménez López, Estela
author2_role author
author
author
dc.subject.none.fl_str_mv Electroforesi capil·lar
Espectrometria de masses
Glicopèptids
Capillary electrophoresis
Mass spectrometry
Glycopeptides
topic Electroforesi capil·lar
Espectrometria de masses
Glicopèptids
Capillary electrophoresis
Mass spectrometry
Glycopeptides
description In this study, several chromatographic sorbents: porous graphitic carbon (PGC), aminopropyl hydrophilic interaction (aminopropyl-HILIC), and phenylboronic acid (PBA) were assessed for the analysis of glycopeptides by on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CEMS). As the PBA sorbent provided the most promising results, a PBA-SPE-CE-MS method was developed for the selective and sensitive preconcentration of glycopeptides from enzymatic digests of glycoproteins. Recombinant human erythropoietin (rhEPO) was selected as the model glycoprotein and subjected to enzymatic digestion with several proteases. The tryptic O126 and N83 glycopeptides from rhEPO were targeted to optimize the methodology. Under the optimized conditions, intraday precision, linearity, limits of detection (LODs), and microcartridge lifetime were evaluated, obtaining improved results compared to that from a previously reported TiO2-SPE-CE-MS method, especially for LODs of N-glycopeptides (up to 500 times lower than by CE-MS and up to 200 times lower than by TiO2-SPE-CE-MS). Moreover, rhEPO Glu-C digests were also analyzed by PBA-SPE-CE-MS to better characterize N24 and N38 glycopeptides. Finally, the established method was used to analyze two rhEPO products (EPOCIM and NeuroEPO plus), demonstrating its applicability in biopharmaceutical analysis. The sensitivity of the proposed PBA-SPE-CEMS method improves the existing CE-MS methodologies for glycopeptide analysis and shows a great potential in glycoprotein analysis to deeply characterize protein glycosites even at low concentrations of the protein digest.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/199374
url https://hdl.handle.net/2445/199374
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1021/acs.jproteome.2c00569
Journal of Proteome Research, 2022, vol. 22, p. 826-836
https://doi.org/10.1021/acs.jproteome.2c00569
dc.rights.none.fl_str_mv cc-by (c) Mancera Arteu, Montserrat et al. , 2022
http://creativecommons.org/licenses/by/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by (c) Mancera Arteu, Montserrat et al. , 2022
http://creativecommons.org/licenses/by/3.0/es/
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 15,300724

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