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

Holtta, Mikko; Minthon, Lennart LU ; Hansson, Oskar LU ; Holmen-Larsson, Jessica; Pike, Ian; Ward, Malcolm; Kuhn, Karsten; Ruetschi, Ulla; Zetterberg, Henrik and Blennow, Kaj, et al. (2015) In Journal of Proteome Research 14(2). p.654-663
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... (More)
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). (Less)
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Contribution to journal
publication status
published
subject
keywords
proteomics, peptidomics, Alzheimer's disease, neurodegenerative disease, biomarker discovery, cerebrospinal fluid, isobaric labeling, quantification, clinical protemics
in
Journal of Proteome Research
volume
14
issue
2
pages
654 - 663
publisher
The American Chemical Society
external identifiers
  • wos:000349276400007
  • scopus:84922680605
ISSN
1535-3893
DOI
10.1021/pr501076j
language
English
LU publication?
yes
id
22429aad-ec5b-4989-a894-2647b058efa9 (old id 5160259)
date added to LUP
2015-04-01 07:40:30
date last changed
2017-11-05 03:17:12
@article{22429aad-ec5b-4989-a894-2647b058efa9,
  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).},
  author       = {Holtta, Mikko and Minthon, Lennart and Hansson, Oskar and Holmen-Larsson, Jessica and Pike, Ian and Ward, Malcolm and Kuhn, Karsten and Ruetschi, Ulla and Zetterberg, Henrik and Blennow, Kaj and Gobom, Johan},
  issn         = {1535-3893},
  keyword      = {proteomics,peptidomics,Alzheimer's disease,neurodegenerative disease,biomarker discovery,cerebrospinal fluid,isobaric labeling,quantification,clinical protemics},
  language     = {eng},
  number       = {2},
  pages        = {654--663},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {An Integrated Workflow for Multiplex CSF Proteomics and Peptidomics-Identification of Candidate Cerebrospinal Fluid Biomarkers of Alzheimer's Disease},
  url          = {http://dx.doi.org/10.1021/pr501076j},
  volume       = {14},
  year         = {2015},
}