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PXD039282

PXD039282 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleSENSITIVE ANALYSIS OF RECOMBINANT HUMAN ERYTHROPOIETIN GLYCOPEPTIDES BY ON-LINE PHENYLBORONIC ACID SOLID-PHASE EXTRACTION CAPILLARY ELECTROPHORESIS-MASS SPECTROMETRY
DescriptionIn 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-CE-MS). As 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 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, repeatabilityintraday precision, linearity, limits of detection (LODs) and microcartridge lifetime were evaluated, obtaining improved results compared to 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 Finally, the PBA-SPE-CE-MS method was applied to the analysis of rhEPO glycopeptides from Glu-C digests to better characterize N24 and N38 glycopeptidessites. Finally, the established method was used to analyze two rhEPO biosimilars (EPOCIM and NeuroEPO plus), demonstrating its applicability in biopharmaceutical analysis. The sensitivity of the proposed PBA-SPE-CE-MS 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 protein digest.
HostingRepositoryPRIDE
AnnounceDate2023-11-14
AnnouncementXMLSubmission_2023-11-14_08:00:37.812.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterMontserrat Mancera Arteu
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
Instrument6220 Time-of-Flight LC/MS
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-01-05 05:55:35ID requested
12023-02-13 02:23:54announced
22023-11-14 08:00:39announced2023-11-14: Updated project metadata.
Publication List
Mancera-Arteu M, Benavente F, Sanz-Nebot V, Gim, é, nez E, Sensitive Analysis of Recombinant Human Erythropoietin Glycopeptides by On-Line Phenylboronic Acid Solid-Phase Extraction Capillary Electrophoresis Mass Spectrometry. J Proteome Res, 22(3):826-836(2023) [pubmed]
Keyword List
submitter keyword: Capillary electrophoresis
glycopeptides
mass spectrometry
in-line solid-phase extraction
on-line solid-phase extraction
phenylboronic acid
Contact List
Estela Giménez López
contact affiliationDepartment of Chemical Engineering and Analytical Chemistry, Institute for Research on Nutrition and Food Safety (INSA·UB), University of Barcelona, Martí i Franquès 1-11, 08028 Barcelona, Spain
contact emailestelagimenez@ub.edu
lab head
Montserrat Mancera Arteu
contact affiliationUniversity of Barcelona
contact emailmontsemancera@ub.edu
dataset submitter
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Dataset FTP location
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