PXD010115-2

PXD010115 is an original dataset announced via ProteomeXchange.

Title	Quantification of the Soluble Receptor of Advanced Glycation End-Products (sRAGE) by LC-MS Without the Use of Affinity Enrichment
Description	The 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.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:45:54.445.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD010115
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Horvatovich Péter
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	Oxidation; Carbamidomethyl
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2018-06-13 07:44:18	ID requested
1	2019-04-09 04:04:54	announced
⏵ 2	2024-10-22 04:46:03	announced	2024-10-22: Updated project metadata.

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]

10.1016/j.aca.2018.09.050;

10.6019/PXD010115;

curator keyword: Biological

submitter keyword: sRAGE, human serum, SCX fractionation, LC-MS/MS

Peter Horvatovich
contact affiliation	Department of Analytical Biochemistry, Groningen Research Institute of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands
contact email	p.l.horvatovich@rug.nl
lab head
Horvatovich Péter
contact affiliation	Faculty of Science and Engineering, Groningen Research Institute of Pharmacy, Analytical Biochemistry, University of Groningen, 9713 AV Groningen, The Netherlands
contact email	p.l.horvatovich@rug.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010115
     1. Label: PRIDE project
     2. Name: Quantification of the Soluble Receptor of Advanced Glycation End-Products (sRAGE) by LC-MS Without the Use of Affinity Enrichment