From Relative to Absolute Quantification of Tryptic Peptides with Tandem Mass Tags: Application to Cerebrospinal Fluid

Authors

  • Loïc Dayon Biomedical Proteomics Group, University of Geneva, CH-1211 Geneva 4
  • Natacha Turck Biomedical Proteomics Group, University of Geneva, CH-1211 Geneva 4
  • Alexander Scherl Biomedical Proteomics Group, University of Geneva, CH-1211 Geneva 4
  • Denis F. Hochstrasser Biomedical Proteomics Group, University of Geneva, CH-1211 Geneva 4; Clinical Proteomics Group, Geneva University Hospitals, CH-1211 Geneva 14
  • Pierre R. Burkhard Department of Neurology, Geneva University Hospitals and Faculty of Medicine, CH-1211 Geneva 14
  • Jean-Charles Sanchez Biomedical Proteomics Group, University of Geneva, CH-1211 Geneva 4

DOI:

https://doi.org/10.2533/chimia.2010.132

Keywords:

Brain disorders, Cerebrospinal fluid, Mass spectrometry, Proteomics, Quantification

Abstract

Quantification is a major task in proteomics. Among the different analytical strategies to enable peptide and protein quantification, tagging with isotopic labels has emerged as a practical, versatile, and efficient alternative. In particular, isobaric labels, such as TMT or iTRAQ, are now widely employed to make relative comparison of the protein amounts in separate biological samples with tandem mass spectrometry (MS/MS). We used herein a shotgun proteomic approach based on labelling with tandem mass tags (TMTs) for the relative quantification of proteins, and the absolute quantification of their tryptic peptides in human cerebrospinal fluid (CSF). First, the comparison of ante- and post-mortem CSF samples was carried out for the discovery of protein marker candidates of brain-damage disorders. Second, tryptic peptides representative of these candidates were measured in CSF using reporter-ion calibration curves. These works highlighted the advantages and limitations of such strategies for quantification purposes in proteomics.

Downloads

Published

2010-03-31

How to Cite

[1]
L. Dayon, N. Turck, A. Scherl, D. F. Hochstrasser, P. R. Burkhard, J.-C. Sanchez, Chimia 2010, 64, 132, DOI: 10.2533/chimia.2010.132.