Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry

In this study is described an on-line titanium dioxide solid-phase extraction capillary electrophoresis-mass spectrometry (TiO2-SPE-CE-MS) method for the analysis of the glycopeptide glycoforms obtained from the tryptic digests of recombinant human erythropoietin (rhEPO). The O126-glycopeptide of rh...

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Autores: Mancera Arteu, Montserrat, Lleshi, Nejsi, Sanz Nebot, María Victoria, Giménez López, Estela, Benavente Moreno, Fernando J. (Julián)
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2020
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/184803
Acceso en línea:https://hdl.handle.net/2445/184803
Access Level:acceso abierto
Palabra clave:Diòxid de titani
Espectrometria de masses
Marcadors bioquímics
Electroforesi capil·lar
Glicoproteïnes
Titanium dioxide
Mass spectrometry
Biochemical markers
Capillary electrophoresis
Glycoproteins
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repository_id_str
spelling Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometryMancera Arteu, MontserratLleshi, NejsiSanz Nebot, María VictoriaGiménez López, EstelaBenavente Moreno, Fernando J. (Julián)Diòxid de titaniEspectrometria de massesMarcadors bioquímicsElectroforesi capil·larGlicoproteïnesTitanium dioxideMass spectrometryBiochemical markersCapillary electrophoresisGlycoproteinsIn this study is described an on-line titanium dioxide solid-phase extraction capillary electrophoresis-mass spectrometry (TiO2-SPE-CE-MS) method for the analysis of the glycopeptide glycoforms obtained from the tryptic digests of recombinant human erythropoietin (rhEPO). The O126-glycopeptide of rhEPO was used to optimize the methodology given its importance in quality control of biopharmaceuticals and doping analysis. Several aspects that affect the selective retention and elution, peak efficiency and electrophoretic separation of the O126 glycoforms were investigated to maximize detection sensitivity while minimizing non-specific retention of peptides. Under the optimized conditions, the microcartridge lifetime was around 10 analyses and repeatability was acceptable (%RSD values of 9-11% and 6-11% for migration times and peak areas, respectively). The method was linear between 0.5-50 mg·L-1 and 10-50 mg·L-1 for O126 glycoforms containing NeuAc and NeuGc, respectively, and limits of detection (LODs) were up to 100 times lower than by CE-MS. Although optimized for O-glycopeptides, the method proved also successful for preconcentration of N83-glycopeptides, without compromising the separation between glycopeptide glycoforms with different number of sialic acids. Tryptic digests of other glycoproteins (i.e. human apolipoprotein CIII (APO-C3) and bovine alpha-1-acid glycoprotein (bAGP)) were also analyzed, demonstrating the applicability to glycopeptides with different glycan composition and nature.Elsevier B.V.2022202220202022info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersion38 p.application/pdfhttps://hdl.handle.net/2445/184803Articles publicats en revistes (Enginyeria Química i Química Analítica)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésVersió postprint del document publicat a: https://doi.org/10.1016/j.talanta.2019.120563Talanta, 2020, vol. 209, num. 120563https://doi.org/10.1016/j.talanta.2019.120563cc-by-nc-nd (c) Elsevier B.V., 2020https://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:2445/1848032026-05-29T05:05:01Z
dc.title.none.fl_str_mv Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
title Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
spellingShingle Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
Mancera Arteu, Montserrat
Diòxid de titani
Espectrometria de masses
Marcadors bioquímics
Electroforesi capil·lar
Glicoproteïnes
Titanium dioxide
Mass spectrometry
Biochemical markers
Capillary electrophoresis
Glycoproteins
title_short Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
title_full Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
title_fullStr Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
title_full_unstemmed Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
title_sort Analysis of glycopeptide biomarkers by on-line TiO2 solid-phase extraction capillary electrophoresis-mass spectrometry
dc.creator.none.fl_str_mv Mancera Arteu, Montserrat
Lleshi, Nejsi
Sanz Nebot, María Victoria
Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
author Mancera Arteu, Montserrat
author_facet Mancera Arteu, Montserrat
Lleshi, Nejsi
Sanz Nebot, María Victoria
Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
author_role author
author2 Lleshi, Nejsi
Sanz Nebot, María Victoria
Giménez López, Estela
Benavente Moreno, Fernando J. (Julián)
author2_role author
author
author
author
dc.subject.none.fl_str_mv Diòxid de titani
Espectrometria de masses
Marcadors bioquímics
Electroforesi capil·lar
Glicoproteïnes
Titanium dioxide
Mass spectrometry
Biochemical markers
Capillary electrophoresis
Glycoproteins
topic Diòxid de titani
Espectrometria de masses
Marcadors bioquímics
Electroforesi capil·lar
Glicoproteïnes
Titanium dioxide
Mass spectrometry
Biochemical markers
Capillary electrophoresis
Glycoproteins
description In this study is described an on-line titanium dioxide solid-phase extraction capillary electrophoresis-mass spectrometry (TiO2-SPE-CE-MS) method for the analysis of the glycopeptide glycoforms obtained from the tryptic digests of recombinant human erythropoietin (rhEPO). The O126-glycopeptide of rhEPO was used to optimize the methodology given its importance in quality control of biopharmaceuticals and doping analysis. Several aspects that affect the selective retention and elution, peak efficiency and electrophoretic separation of the O126 glycoforms were investigated to maximize detection sensitivity while minimizing non-specific retention of peptides. Under the optimized conditions, the microcartridge lifetime was around 10 analyses and repeatability was acceptable (%RSD values of 9-11% and 6-11% for migration times and peak areas, respectively). The method was linear between 0.5-50 mg·L-1 and 10-50 mg·L-1 for O126 glycoforms containing NeuAc and NeuGc, respectively, and limits of detection (LODs) were up to 100 times lower than by CE-MS. Although optimized for O-glycopeptides, the method proved also successful for preconcentration of N83-glycopeptides, without compromising the separation between glycopeptide glycoforms with different number of sialic acids. Tryptic digests of other glycoproteins (i.e. human apolipoprotein CIII (APO-C3) and bovine alpha-1-acid glycoprotein (bAGP)) were also analyzed, demonstrating the applicability to glycopeptides with different glycan composition and nature.
publishDate 2020
dc.date.none.fl_str_mv 2020
2022
2022
2022
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/2445/184803
url https://hdl.handle.net/2445/184803
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/10.1016/j.talanta.2019.120563
Talanta, 2020, vol. 209, num. 120563
https://doi.org/10.1016/j.talanta.2019.120563
dc.rights.none.fl_str_mv cc-by-nc-nd (c) Elsevier B.V., 2020
https://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by-nc-nd (c) Elsevier B.V., 2020
https://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 38 p.
application/pdf
dc.publisher.none.fl_str_mv Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv Articles publicats en revistes (Enginyeria Química i Química Analítica)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869412856031608832
score 15,811543

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