Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis

NeuroEPO plus is a recently developed recombinant human erythropoietin (rhEPO) without erythropoietic activity and shorter plasma half-life due to its low sialic acid content. This novel rhEPO product is under investigation as therapeutic protein in the treatment of neurodegenerative diseases owing...

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Detalles Bibliográficos
Autores: García-Artalejo, Judey A., Mancera Arteu, Montserrat, Sanz Nebot, María Victoria, Rodríguez, Teresita, Giménez López, Estela
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
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:2445/187406
Acceso en línea:https://hdl.handle.net/2445/187406
Access Level:acceso abierto
Palabra clave:Espectrometria de masses
Glicoproteïnes
Polisacàrids
Mass spectrometry
Glycoproteins
Polysaccharides
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spelling Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysisGarcía-Artalejo, Judey A.Mancera Arteu, MontserratSanz Nebot, María VictoriaRodríguez, TeresitaGiménez López, EstelaEspectrometria de massesGlicoproteïnesPolisacàridsMass spectrometryGlycoproteinsPolysaccharidesNeuroEPO plus is a recently developed recombinant human erythropoietin (rhEPO) without erythropoietic activity and shorter plasma half-life due to its low sialic acid content. This novel rhEPO product is under investigation as therapeutic protein in the treatment of neurodegenerative diseases owing to its neuroprotective and neurodegenerative properties. In this study, an in-depth characterization of NeuroEPO plus N-glycans was performed by a glycan isotope [12C6]/[13C6] coded aniline labeling strategy followed by capillary zwitterionic hydrophilic interaction liquid chromatography-mass spectrometry (CapZIC-HILIC-MS). A superior amount of low sialylated glycans and less branched structures were detected in NeuroEPO plus compare to other commercial rhEPOs. At the intact glycoprotein level, NeuroEPO plus glycoforms were separated by capillary zone electrophoresis with ultraviolet detection (CE-UV), optimizing the composition and pH of the separation electrolyte. Moreover, an isoelectric focusing polyacrylamide gel electrophoresis (IEF-PAGE) method was also optimized for the simultaneous analysis of this basic rhEPO and conventional acidic rhEPO products. The proposed glycomic and intact glycoprotein methods provide a robust and reliable analytical platform for NeuroEPO plus characterization and for its future implementation as biopharmaceutical in neurodegenerative diseases.Elsevier B.V.2022202220222022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/2445/187406Articles 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)InglésReproducció del document publicat a: https://doi.org/10.1016/j.jpba.2022.114686Journal of Pharmaceutical and Biomedical Analysis, 2022, vol. 213, p. 114686https://doi.org/10.1016/j.jpba.2022.114686cc-by-nc-nd (c) García-Artalejo, Judey A., et al., 2022https://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:2445/1874062026-05-29T05:05:01Z
dc.title.none.fl_str_mv Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
title Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
spellingShingle Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
García-Artalejo, Judey A.
Espectrometria de masses
Glicoproteïnes
Polisacàrids
Mass spectrometry
Glycoproteins
Polysaccharides
title_short Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
title_full Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
title_fullStr Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
title_full_unstemmed Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
title_sort Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis
dc.creator.none.fl_str_mv García-Artalejo, Judey A.
Mancera Arteu, Montserrat
Sanz Nebot, María Victoria
Rodríguez, Teresita
Giménez López, Estela
author García-Artalejo, Judey A.
author_facet García-Artalejo, Judey A.
Mancera Arteu, Montserrat
Sanz Nebot, María Victoria
Rodríguez, Teresita
Giménez López, Estela
author_role author
author2 Mancera Arteu, Montserrat
Sanz Nebot, María Victoria
Rodríguez, Teresita
Giménez López, Estela
author2_role author
author
author
author
dc.subject.none.fl_str_mv Espectrometria de masses
Glicoproteïnes
Polisacàrids
Mass spectrometry
Glycoproteins
Polysaccharides
topic Espectrometria de masses
Glicoproteïnes
Polisacàrids
Mass spectrometry
Glycoproteins
Polysaccharides
description NeuroEPO plus is a recently developed recombinant human erythropoietin (rhEPO) without erythropoietic activity and shorter plasma half-life due to its low sialic acid content. This novel rhEPO product is under investigation as therapeutic protein in the treatment of neurodegenerative diseases owing to its neuroprotective and neurodegenerative properties. In this study, an in-depth characterization of NeuroEPO plus N-glycans was performed by a glycan isotope [12C6]/[13C6] coded aniline labeling strategy followed by capillary zwitterionic hydrophilic interaction liquid chromatography-mass spectrometry (CapZIC-HILIC-MS). A superior amount of low sialylated glycans and less branched structures were detected in NeuroEPO plus compare to other commercial rhEPOs. At the intact glycoprotein level, NeuroEPO plus glycoforms were separated by capillary zone electrophoresis with ultraviolet detection (CE-UV), optimizing the composition and pH of the separation electrolyte. Moreover, an isoelectric focusing polyacrylamide gel electrophoresis (IEF-PAGE) method was also optimized for the simultaneous analysis of this basic rhEPO and conventional acidic rhEPO products. The proposed glycomic and intact glycoprotein methods provide a robust and reliable analytical platform for NeuroEPO plus characterization and for its future implementation as biopharmaceutical in neurodegenerative diseases.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
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/2445/187406
url https://hdl.handle.net/2445/187406
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/10.1016/j.jpba.2022.114686
Journal of Pharmaceutical and Biomedical Analysis, 2022, vol. 213, p. 114686
https://doi.org/10.1016/j.jpba.2022.114686
dc.rights.none.fl_str_mv cc-by-nc-nd (c) García-Artalejo, Judey A., et al., 2022
https://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by-nc-nd (c) García-Artalejo, Judey A., et al., 2022
https://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
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
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score 15,812429

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