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Giuseppina Maccarrone Isabel Birg Eva Malisch Marcus C. Rosenhagen Claudia Ditzen John A. Chakel Friedrich Mandel Andreas Reimann Can-Carlo Doertbudak Katrin Haegler Florian Holsboer Christoph W. Turck PhD 《Clinical proteomics》2004,1(3-4):333-364
The identification of disease markers in human body fluids requires an extensive and thorough analysis of its protein constituents.
In the present study, we have extended our analysis of the human cerebrospinal fluid (CSF) proteome using protein prefractional
followed by shotgun mass spectrometry. After the removal of abundant protein components from the mixture with the help of
immunodepletion affinity chromatography, we used either anion exchange chromatography or sodium dodecyl sulfate-polyacrylamide
gel electrophoresis (SDS-PAGE) to further subfractionate the proteins present in CSFs. Each protein subfraction was enzyme
digested and analyzed by tandem mass spectrometry and the resulting data evaluated using the Spectrum Mill software. Different
subfractionation methods resulted in the identification of a grant total of 259 proteins in CSF from a patient with normal
pressure hydrocephalus. The greatest number of protein, 240 in total, were identified after prefractionating the CSF proteins
by immunodepletion and SDS-PAGE. Immuno-depletion combined with anion exchange fractionation resulted in 112 proteins and
74 proteins were found when only immunodepletion of the CSF samples was carried out. All methods used showed a significant
increase in the number of identified proteins as compared with nondepleted and unfractionated CSF sample analysis, which yielded
only 38 protein identifications. The present work establishes a platform for future studies aimed at a detailed comparative
proteome analysis of CSFs from different groups of patients suffering from various psychiatric and neurological disorders. 相似文献
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