An Integrated Workflow for Multiplex CSF Proteomics and Peptidomics-Identification of Candidate Cerebrospinal Fluid Biomarkers of Alzheimer's Disease

Research output: Contribution to journalArticle

Abstract

Many disease processes in the brain are reflected in the protein composition of the cerebrospinal fluid (CSF). In addition to proteins, CSF also contains a large number of endogenous peptides whose potential as disease biomarkers largely remains to be explored. We have developed a novel workflow in which multiplex isobaric labeling is used for simultaneous quantification of endogenous CSF peptides and proteins by liquid chromatography coupled with mass spectrometry. After the labeling of CSF samples, endogenous peptides are separated from proteins by ultrafiltration. The proteins retained on the filters are trypsinized, and the tryptic peptides are collected separately. We evaluated this technique in a comparative pilot study of CSF peptide and protein profiles in eight patients with Alzheimer's disease (AD) and eight nondemented controls. We identified several differences between the AD and control group among endogenous peptides derived from proteins known to be associated with AD, including neurosecretory protein VGF (ratios AD/controls 0.45-0.81), integral membrane protein 2B (ratios AD/controls 0.72-0.84), and metallothionein-3 (ratios AD/controls 0.51-0.61). Analysis of tryptic peptides identified several proteins that were altered in the AD group, some of which have previously been reported as changed in AD, for example, VGF (ratio AD/controls 0.70).

Details

Authors
  • Mikko Holtta
  • Lennart Minthon
  • Oskar Hansson
  • Jessica Holmen-Larsson
  • Ian Pike
  • Malcolm Ward
  • Karsten Kuhn
  • Ulla Ruetschi
  • Henrik Zetterberg
  • Kaj Blennow
  • Johan Gobom
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Neurology

Keywords

  • proteomics, peptidomics, Alzheimer's disease, neurodegenerative disease, biomarker discovery, cerebrospinal fluid, isobaric labeling, quantification, clinical protemics
Original languageEnglish
Pages (from-to)654-663
JournalJournal of Proteome Research
Volume14
Issue number2
Publication statusPublished - 2015
Publication categoryResearch
Peer-reviewedYes