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iTRAQ-Based LC-LC MALDI TOF/TOF MS Quantitative Analysis of Membrane Proteins from Human Glioma

  • Uroš RajčevićEmail author
Protocol
Part of the Neuromethods book series (NM, volume 57)

Abstract

Various proteomic approaches are being applied in brain tumor proteomics with regard to targeted proteins of interest, to discover phenotype specific markers which could facilitate diagnosis as well as potential antitumor drug targets. iTRAQ technology is a multiplexing protein quantitation strategy that provides relative and absolute measurements of protein abundance in complex mixtures based on the differential labeling of the proteins. Combined with membrane enrichment methodology, separation of the labeled peptides by two-dimensional liquid chromatography linked to tandem mass spectrometry and adequate data-mining, it can provide an excellent tool in search for novel isoform- and species-specific biomarkers and drug targets in various biotechnological and biomedical applications.Glioblastoma (GBM) is the most frequent primary brain tumor diagnosed in adults and remains one of the most lethal forms of human cancer. No biomarkers can distinguish different cell populations within GBMs or predict the potential of low grade gliomas to develop into malignant angiogenic gliomas. In our study, we have used iTRAQ-based technology to search for novel biomarkes of GBM.

Key words

iTRAQ Glioblastoma Biomarkers Glioma invasion Angiogenesis Subproteome Quantitative mass spectrometry 2D-LC 

Notes

Acknowledgements

The work presented in this chapter was supported by EU FP6 Integrated project grant “Angiotargeting” (contract number 504743), by CRP-Santé, the Research Ministry (MCESR) in Luxembourg and by grant AFR-PDR-08-007 from the FNR, Luxembourg. The author is grateful to Dr. S.P. Niclou and Prof. R. Bjerkvig NorLux Neuro-Oncology laboratory, CRP-Santé, Luxembourg and Department of Biomedicine, University in Bergen, Norway for their invaluable support with this project and critical review of the manuscript. Drs. C. R. Jimenez and J. Knol, Oncoproteomics Laboratory, VU University Medical Center in Amsterdam, The Netherlands are kindly acknowledged for their help.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Norlux Neuro-Oncology LaboratoryCRP-SantéStrassenLuxembourg
  2. 2.Department of Genetic Toxicology and Cancer BiologyNational Institute of BiologyLjubljanaSlovenia

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