Saliva as a source of new phosphopeptide biomarkers: Development of a comprehensive analytical method based on shotgun peptidomics

TY - JOUR

T1 - Saliva as a source of new phosphopeptide biomarkers

T2 - Development of a comprehensive analytical method based on shotgun peptidomics

AU - La Barbera, Giorgia

AU - Capriotti, Anna Laura

AU - Cavaliere, Chiara

AU - Ferraris, Francesca

AU - Montone, Carmela Maria

AU - Piovesana, Susy

AU - Zenezini Chiozzi, Riccardo

AU - Laganà, Aldo

PY - 2018/6/1

Y1 - 2018/6/1

N2 - The paper describes the development of an enrichment method for the analysis of the endogenous phosphopeptides in saliva. The method was based on magnetic solid phase extraction by a magnetic graphitized carbon black-TiO2 composite material and was developed considering different saliva pre-treatments, namely C18 solid phase extraction for purification, direct dilution in loading buffer or acetonitrile precipitation. The method was based on a shotgun proteomics workflow and the enriched peptide mixture was analysed by nanoHPLC and high resolution tandem mass spectrometry. Acetonitrile precipitation provided the best results, with up to 165 endogenous phosphopeptides identified in saliva samples from healthy individuals. The physico-chemical features of the identified endogenous phosphopeptides indicated that such peptides were large, hydrophilic and basic.

AB - The paper describes the development of an enrichment method for the analysis of the endogenous phosphopeptides in saliva. The method was based on magnetic solid phase extraction by a magnetic graphitized carbon black-TiO2 composite material and was developed considering different saliva pre-treatments, namely C18 solid phase extraction for purification, direct dilution in loading buffer or acetonitrile precipitation. The method was based on a shotgun proteomics workflow and the enriched peptide mixture was analysed by nanoHPLC and high resolution tandem mass spectrometry. Acetonitrile precipitation provided the best results, with up to 165 endogenous phosphopeptides identified in saliva samples from healthy individuals. The physico-chemical features of the identified endogenous phosphopeptides indicated that such peptides were large, hydrophilic and basic.

KW - Endogenous saliva phosphopeptides

KW - Magnetic solid phase extraction

KW - Metal oxide affinity chromatography

KW - NanoHPLC-M/MS

KW - Phosphopeptide enrichment

KW - Phosphoproteomics

UR - http://www.scopus.com/inward/record.url?scp=85042518590&partnerID=8YFLogxK

U2 - 10.1016/j.talanta.2018.02.085

DO - 10.1016/j.talanta.2018.02.085

M3 - Journal article

C2 - 29567172

AN - SCOPUS:85042518590

SN - 0039-9140

VL - 183

SP - 245

EP - 249

JO - Talanta

JF - Talanta

ER -