title: Lumbar cerebrospinal fluid proteome in multiple sclerosis: characterization by ultrafiltration, liquid chromatography, and mass spectrometry.
abstract: Neurological diseases, including multiple sclerosis (M.S.), often provoke changes in the functioning of the endothelial and epithelial brain barriers and give rise to disease-associated alterations of the cerebrospinal fluid (CSF) proteome. In the present study, pooled and ultrafiltered CSF of M.S. and non-M.S. patients was digested with trypsin and analyzed by off-line strong cation-exchange chromatography (SCX) coupled to on-line reversed-phase LC-ESI-MS/MS. In an alternative approach, the trypsin-treated subproteomes were analyzed directly by LC-ESI-MS/MS and gas-phase fractionation in the mass spectrometer. Taken together, both proteomic approaches in combination with a three-step evaluation process including the search engines Sequest and Mascot, and the validation software Scaffold, resulted in the identification of 148 proteins. Sixty proteins were identified in CSF for the first time by mass spectrometry. For validation purposes, the concentration of cystatin A was determined in individual CSF and serum samples of M.S. and non-M.S. patients using ELISA.
authors.affiliation: Biomedical Research Institute, Hasselt University, and School of Life Sciences, transnationale Universiteit Limburg, Diepenbeek, Belgium.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Noben
authors.forename: Debora
authors.initials: D
authors.lastname: Dumont
authors.forename: Natalia
authors.initials: N
authors.lastname: Kwasnikowska
authors.forename: Peter
authors.initials: P
authors.lastname: Verhaert
authors.forename: Veerle
authors.initials: V
authors.lastname: Somers
authors.forename: Raymond
authors.initials: R
authors.lastname: Hupperts
authors.forename: Piet
authors.initials: P
authors.lastname: Stinissen
authors.forename: Johan
authors.initials: J
authors.lastname: Robben
mesh_terms: D000328:Adult; D000368:Aged; D016022:Case-Control Studies; D002555:Cerebrospinal Fluid / Q000737:chemistry*; D002853:Chromatography, Liquid; D005260:Female; D006801:Humans; D008159:Lumbar Vertebrae / Q000033:anatomy & histology; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D014462:Ultrafiltration
chemical_list: D020543:Proteome