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Comparison of blood serum peptide enrichment methods by Tricine SDS-PAGE and mass spectrometry

Biosa, Grazia and Addis, Maria Filippa and Tanca, Alessandro and Pisanu, Salvatore and Roggio, Tonina and Uzzau, Sergio and Pagnozzi, Daniela (2011) Comparison of blood serum peptide enrichment methods by Tricine SDS-PAGE and mass spectrometry. Journal of Proteomics, Vol. 75 (1), p. 93-99. ISSN 1874-3919. eISSN 1876-7737. Article.

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DOI: 10.1016/j.jprot.2011.06.025

Abstract

Characterisation of blood serum peptides can provide valuable information on physiological and pathological processes. However, the analysis of raw serum samples by MS results in the identification of a limited number of peptides. In order to improve sensitivity, many peptide enrichment methods have been proposed during the last ten years. Here, we present a comparison of fractionation methods aimed to simplify analysis of small proteins and peptides in blood serum, one of the most promising sources of putative biomarkers. Specifically, three methods based on ultrafiltration, differential precipitation, and peptide ligand libraries (ProteoMiner) were evaluated for the enrichment of peptides and low molecular weight proteins, as demonstrated by Tricine SDS-PAGE and subsequent LC-MS/MS (GeLC-MS/MS). As a result, differential solubilisation (DS) allowed the identification of the highest number of peptides. Moreover, the DS method enabled also the quantitative comparison of samples, producing fundamental information in biomarker discovery approaches.

Item Type:Article
ID Code:6379
Status:Published
Refereed:Yes
Uncontrolled Keywords:Peptidome, mass spectrometry, enrichment, method comparison
Subjects:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
Publisher:Elsevier
ISSN:1874-3919
eISSN:1876-7737
Copyright Holders:© 2011 Elsevier
Deposited On:23 Aug 2011 10:14

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