Standardized peptidome profiling of human cerebrospinal fluid by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Mathias Bruegel*, Mathis Planert, Sven Baumann, Almut Focke, Florian Then Bergh, Alexander Leichtle, Uta Ceglarek, Joachim Thiery, Georg Martin Fiedler

*Korrespondierende/r Autor/-in für diese Arbeit
    26 Zitate (Scopus)

    Abstract

    Peptidome profiling of human cerebrospinal fluid (CSF) is a promising tool to identify novel disease-associated biomarkers. Our aim was to develop a standardized protocol for reproducible peptidome profiling of CSF using magnetic bead (MB) separation followed by MALDI-TOF MS. Peptidome fractionation and profiling of CSF were performed using MBs with different surface functionalities. We investigated exogenous variables (storage conditions, freeze-thaw-cycles) and endogenous interferences (albumin, immunoglobulin, blood, leukocytes) in pooled CSF samples. We detected approximately 500 signals with an S/N ratio > 10 and an overlap frequency of about 40% in non-pathological CSF. Within- and between-day imprecisions in relative signal intensities ranged from 3 to 28% and 7 to 47%, respectively. CSF storage at room temperature for up to 6 h and at 4 °C for up to 3 days did not significantly influence the mass spectra. Consecutive freeze-thaw-cycles significantly affected the mass spectra. High albumin and immunoglobulin content altered the CSF preparation using MB-HIC C8 beads. Blood contamination showed no effect on mass spectra up to a hemoglobin concentration of 0.075 μmol/L. The presence of leukocytes up to a cell number of 30 Mpt/L did not affect mass spectra. Our reliable pretreatment protocol allows standardization of preanalytical modalities and thereby enables reproducible peptidome profiling of human CSF using MB separation followed by MALDI-TOF MS.

    OriginalspracheEnglisch
    ZeitschriftJournal of Proteomics
    Jahrgang72
    Ausgabenummer4
    Seiten (von - bis)608-615
    Seitenumfang8
    ISSN1874-3919
    DOIs
    PublikationsstatusVeröffentlicht - 02.05.2009

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    • Forschungsschwerpunkt: Biomedizintechnik

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