Title: Pre-analytical factors in clinical proteomics investigations: Impact of ex vivo protein modifications for multiple sclerosis biomarker discovery
Authors: Pieragostino, Damiana
Petrucci, Francesca
Del Boccio, Piero
Mantini, Dante
Lugaresi, Alessandra
Tiberio, Sara
Onofrj, Marco
Gambi, Domenico
Sacchetta, Paolo
Di Ilio, Carmine
Federici, Giorgio
Urbani, Andrea # ×
Issue Date: Jan-2010
Publisher: Elsevier BV
Series Title: Journal of Proteomics vol:73 issue:3 pages:579-592
Abstract: Serum proteome investigations have raised an incredible interest in the research of novel molecular biomarker, nevertheless few of the proposed evidences have been translated to the clinical practice. One of the limiting factors has been the lack of generally accepted guidelines for clinical proteomics studies and the lack of a robust analytical and pre-analytical ground for the proposed classification models. Pre-analytical issues may results in a deep impact for biomarker discovery campaign. In this study we present a systematic evaluation of sample storage and sampling conditions for clinical proteomics investigations. We have developed and validated a linear MALDI-TOF-MS protein profiling method to explore the low protein molecular weight region (5-20kDa) of serum samples. Data normalization and processing was performed using optimise peak detection routine (LIMPIC) able to describe each group under investigation. Data were acquired either from healthy volunteers and from multiple sclerosis patients in order to highlight ex vivo protein profile alteration related to different physio-pathological conditions. Our data showed critical conditions for serum protein profiles depending on storage times and temperatures: 23 degrees C, 4 degrees C, -20 degrees C and -80 degrees C. We demonstrated that upon a -20 degrees C short term storage, characteristic degradation profiles are associated with different clinical groups. Protein signals were further identified after preparative HPLC separation by peptide sequencing on a nanoLC-Q-TOF TANDEM mass spectrometer. Apolipoprotein A-IV and complement C3 protein fragments, transthyretin and the oxidized isoforms in different apolipoprotein species represent the major molecular features of such a degradation pattern.
ISSN: 1874-3919
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Research Group Neurophysiology
× corresponding author
# (joint) last author

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