Journal
PROTEOMICS
Volume 8, Issue 20, Pages 4357-4366Publisher
WILEY
DOI: 10.1002/pmic.200800375
Keywords
2-D DIGE; Alzheimer's disease; Cerebrospinal fluid; Creutzfeldt-Jakob disease; Lewy-body dementia
Funding
- Landesstiftung Baden-Wurttemberg [P-LS-Prot/42]
- European Commission
- German Ministry of Science and Technology
- German Ministry of Health
- Research Center for Molecular Physiology of the Brain (CMPB)
- Deutsche Forschungsgemeinschaft (DFG)
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So far only the detection of 14-3-3 proteins in cerebrospinal fluid (CSF) is included in the diagnostic criteria for sporadic Creutzfeldt-jakob disease (sCJD). However, this assay cannot be used for screening because of the high rate of false positive results in sCJD, and often negative results in variant CJD. To facilitate the differential diagnosis of CJD, we applied 2-D differential gel-electrophoresis (2-D DIGE) as a quantitative proteomic screening system for CSF proteins. We compared 36 patients suffering from sCJD with 30 patients suffering from other neurodegenerative diseases. Sample preparation was optimized in consideration of the fact that CSF is composed of blood- and brain-derived proteins, and an improved 2-D DIGE protocol was established. Using this method in combination with protein identification by MALDI-TOF-MS, several known surrogate markers of sCJD like 14-3-3 protein, neuron-specific enolase, and lactate dehydrogenase were readily identified. Moreover, a not yet identified protein with an approximate molecular mass of 85 kDa was found as marker for sCJD with high diagnostic specificity and sensitivity. We conclude that our proteomic approach is useful to differentiate CJD from other neurodegenerative diseases and expect that CSF-optimized 2-D DIGE will find broad application in the search for other brain derived proteins in CSF.
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