Proteomic and Mass Spectrometry Technologies for Biomarker Discovery - Proteomic and Metabolomic Approaches to Biomarker Discovery - page 17

CHAPTER
2

Proteomic and Mass Spectrometry
Technolo gies for Biomarker Discovery

Andrei P. Drabovich 1 , Maria P. Pavlou 2 , Ihor Batruch 1 ,
Eleftherios P. Diamandis 1e4
1 Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, ON, Canada,
2 Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada,
3 Department of Clinical Biochemistry, University Health Network, Toronto, ON, Canada,
4 Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON, Canada

OUTLINE

Introduction 18 Label-Free Quanti cation 27

Metabolic and Enzymatic Labeling 28

Protein Biomarker Discovery and
Development Pipeline

Chemical Labeling 29

18

Selected Reaction Monitoring Assays 29

Proteomic Samples 20 Separation and Enrichment Strategies for

Quanti cation of Low-Abundance

Protein Identi cation by Mass Spectrometry 22

Protein Digestion

Proteins 30

23

Protein and Peptide Separation Techniques 23 Biomarker Veri cation 30

Protein and Peptide Ionization Techniques 23

Biomarker Validation 31

Mass Spectrometry Instrumentation 24

Deconvolution and Database Search of
Tandem Mass Spectra

Limitations of Mass Spectrometry for Protein
Biomarker Discovery 25 32

Post-Translational Modi cations as Disease Conclusions and Future Outlook: Integrated

Biomarker Discovery Platform Biomarkers 25 32

Protein Quanti cation by Mass Spectrometry 27 References 33