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Title: ENRICHMENT OF AMYLOID-beta-PEPTIDES FROM CEREBROSPINAL FLUID USING SINGLE SOLID PHASE EXTRACTION BEADS AND SUBSEQUENT MASS SPECTROMETRY ANALYSIS
Authors: Bertinetti, O.
Singer, K.
Bertinetti, D.
Hanke, S.
Boerjan, Bart
Reuner, K. H.
Herberg, F. W.
Issue Date: May-2011
Publisher: Walter de Gruyter
Host Document: Clinical Chemistry and Laboratory Medicine vol:49 pages:S772-S772
Conference: -
Abstract: Background. Clinical diagnostics of Alzheimer‘s disease (AD) focuses on the determination of relative ratios of amyloid-β-peptides (Aβ-peptides) Aβ1-42 and Aβ1-40 in cerebrospinal fluid (CSF). Enrichment of such low abundance peptides on solid phase extraction (SPE) beads in combination with mass spectrometry (MS) provides a technique to detect biomarkers in body fluids.
Methods. Miniaturisation of the SPE technique allowed capturing and subsequent analysis of the secreted Aβ-peptides from body fluids. Salt load and high abandoned liquor proteins were depleted while simultaneously Aβ-peptides where enriched from cerebrospinal fluid on one single SPE bead. Single SPE beads were transferred to a matrix-assisted laser desorption/ionization (MALDI) target and peptides were directly eluted with 2,5-Dihydroxyacetophenone matrix and subsequently analysed on a Bruker Ultraflex MALDI-TOF/TOF with high confidence. CSF from non-demented controls spiked with different Aβ-peptides was tested against AD patient liquor.
Results. Spiked CSF samples allowed an efficient enrichment procedure with subsequent MALDI analysis within minutes. Aβ-peptide concentrations comparably with literature values (< 1µM) could be detected in sample volumes < 50µl. Peptides from patients with high unspecific protein back round (blood brain barrier defects) could still be detected. AD patient samples were tested, peak intensities/areas were determined and compared with the clinical results.
Conclusions. The single bead technique enabled a fast and sensitive analysis of Aβ-peptides in low sample volume even against high protein background.
ISSN: 1434-6621
Publication status: published
KU Leuven publication type: IMa
Appears in Collections:Animal Physiology and Neurobiology Section - miscellaneous

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