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PXD010115

PXD010115 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleQuantification of the Soluble Receptor of Advanced Glycation End-Products (sRAGE) by LC-MS Without the Use of Affinity Enrichment
DescriptionThe study of low abundant proteins contributes to increasing our knowledge about (patho)physiological processes of ageing and disease onset and development and may lead to the identification and clinical application of disease markers. However, studying these proteins is challenging as high-abundant proteins complicate their analysis. Antibodies are often used to enrich proteins from biological matrices prior to their analysis, though antibody-free approaches have been described for some proteins as well. Here we report an antibody-free workflow on the basis of strong cation exchange (SCX) enrichment and liquid chromatography-mass spectrometry (LC-MS) for quantification of the soluble Receptor of Advanced Glycation End-products (sRAGE), a promising biomarker in chronic obstructive pulmonary disease (COPD). sRAGE was quantified in serum at clinically relevant low to sub ng/mL levels. The method was validated according to U.S. Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines and was compared to an antibody-based LC-MS sRAGE method. The SCX-based method builds upon on the bipolar charge distribution of sRAGE, which has a highly basic N-terminal part and an acidic C-terminal part resulting in an overall neutral isoelectric point (pI). The highly basic N-terminal part (pIcalculated = 10.3) allowed sRAGE to be enriched by SCX at pH 10, a pH at which most serum proteins do not bind. This study shows that ion exchange-based enrichment is a viable approach for the LC-MS analysis of several low abundant proteins following a thorough analysis of their physical-chemical properties.
HostingRepositoryPRIDE
AnnounceDate2019-04-09
AnnouncementXMLSubmission_2019-04-09_04:04:53.xml
DigitalObjectIdentifierhttp://dx.doi.org/10.6019/PXD010115
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterHorvatovich Péter
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListOxidation; Carbamidomethyl
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02018-06-13 07:44:18ID requested
12019-04-09 04:04:54announced
Publication List
Klont F, Joosten MR, Ten Hacken NHT, Horvatovich P, Bischoff R, Quantification of the soluble Receptor of Advanced Glycation End-Products (sRAGE) by LC-MS after enrichment by strong cation exchange (SCX) solid-phase extraction (SPE) at the protein level. Anal Chim Acta, 1043():45-51(2018) [pubmed]
Keyword List
curator keyword: Biological
submitter keyword: sRAGE, SCX fractionation, LC-MS/MS, human serum
Contact List
Peter Horvatovich
contact affiliationDepartment of Analytical Biochemistry, Groningen Research Institute of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands
contact emailp.l.horvatovich@rug.nl
lab head
Horvatovich Péter
contact affiliationFaculty of Mathematics and Natural Sciences, Department of Pharmacy, Analytical Biochemistry, University of Groningen, 9713 AV Groningen, The Netherlands
contact emailp.l.horvatovich@rug.nl
dataset submitter
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