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http://hdl.handle.net/1942/1428
Title: | Lumbar cerebrospinal fluid proteome in multiple sclerosis: Characterization by ultrafiltration, liquid chromatography, and mass spectrometry | Authors: | NOBEN, Jean-Paul DUMONT, Debora KWASNIKOWSKA, Natalia VERHAERT, Peter SOMERS, Veerle Hupperts, Raymond STINISSEN, Piet ROBBEN, Johan |
Issue Date: | 2006 | Publisher: | AMER CHEMICAL SOC | Source: | JOURNAL OF PROTEOME RESEARCH, 5(7). p. 1647-1657 | 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. | Keywords: | centrifugal ultrafiltration; cerebrospinal fluid; cystatin A; gas phase; fractionation; liquid chromatography; mass spectrometry; multiple; sclerosis; proteomics; validation; 2-DIMENSIONAL GEL-ELECTROPHORESIS; DIAZEPAM-BINDING INHIBITOR; CREUTZFELDT-JAKOB-DISEASE; CYSTATIN-C; SUBARACHNOID HEMORRHAGE; ALZHEIMERS-DISEASE; CREATINE-KINASE; SAMPLE PREPARATION; STATISTICAL-MODEL; NERVOUS-SYSTEM | Document URI: | http://hdl.handle.net/1942/1428 | ISSN: | 1535-3893 | e-ISSN: | 1535-3907 | DOI: | 10.1021/pr0504788 | ISI #: | 000238838500016 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2007 |
Appears in Collections: | Research publications |
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Noben2006.pdf | Peer-reviewed author version | 211.31 kB | Adobe PDF | View/Open |
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