Open Access

In-depth analysis of the human CSF proteome using protein prefractionation

  • Giuseppina Maccarrone1,
  • Isabel Birg1,
  • Eva Malisch1,
  • Marcus C. Rosenhagen1,
  • Claudia Ditzen1,
  • John A. Chakel2,
  • Friedrich Mandel3,
  • Andreas Reimann3,
  • Can-Carlo Doertbudak1,
  • Katrin Haegler1,
  • Florian Holsboer1 and
  • Christoph W. Turck1Email author
Clinical Proteomics1:13104333

https://doi.org/10.1385/CP:1:3-4:333

Abstract

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.

Key Words

Brain disease cerebrospinal fluid proteome protein prefractionation

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