1. Pre- and Post-Analytical Factors in Biomarker Discovery
Frank Klont, Peter Horvatovich, Natalia Govorukhina, and Rainer Bischoff
2. Pre-Fractionation of Non-Circulating Biological Fluids to Improve Discovery of Clinically Relevant Protein Biomarkers
Annarita Farina
3. Serum Exosome Isolation by Size-Exclusion Chromatography for the Discovery and Validation of Preeclampsia-Associated Biomarkers
Rosana Navajas, Fernando J. Corrales, and Alberto Paradela
4. Protein Biomarker Discovery Using Human Blood Plasma Microparticles
Raghda Saad Zaghloul Taleb, Pacint Moez, Doreen Younan, Martin Eisenacher, Matthias Tenbusch, Barbara Sitek, and Thilo Bracht
5. A Standardized and Reproducible Proteomics Protocol for Bottom-Up Quantitative Analysis of Protein Samples Using SP3 and Mass Spectrometry
Christopher S. Hughes, Poul H. Sorensen, and Gregg B. Morin
6. Analyzing Cerebrospinal Fluid Proteomes to Characterize Central Nervous System Disorders: A Highly Automated Mass Spectrometry-Based Pipeline for Biomarker Discovery
Antonio Núñez Galindo, Charlotte Macron, Ornella Cominetti, and Loïc Dayon
7. Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies
Armin Schniers, Yvonne Pasing, and Terkel Hansen
8. Tube-Gel: A Fast and Effective Sample Preparation Method for High-Throughput Quantitative Proteomics
Leslie Muller, Luc Fornecker, Sarah Cianferani, and Christine Carapito
9. Protein Biomarker Discovery in Non-Depleted Serum by Spectral Library-Based Data-Independent Acquisition Mass Spectrometry
Alexandra Kraut, Mathilde Louwagie, Christophe Bruley, Christophe Masselon, Yohann Couté, Virginie Brun, and Anne-Marie Hesse
10. Discovering Protein Biomarkers from Clinical Peripheral Blood Mononuclear Cells Using Data-Independent Acquisition Mass Spectrometry
Xin Ku and Wei Yan
11. Intact Protein Analysis by LC-MS for Characterizing Biomarkers in Cerebrospinal Fluid
Jérôme Vialaret, Sylvain Lehmann, and Christophe Hirtz
12. Detection of Proteoforms Using Top-Down Mass Spectrometry and Diagnostic Ions
Didia Coelho Graça, Ralf Hartmer, Wolfgang Jabs, Alexander Scherl, Lorella Clerici, Kaveh Samii, Yury O. Tsybin, Denis Hochstrasser, and Pierre Lescuyer
13. Development of a Highly Multiplexed SRM Assay for Biomarker Discovery in Formalin-Fixed Paraffin-Embedded Tissues
Carine Steiner, Pierre Lescuyer, Jean-Christophe Tille, Paul Cutler, and Axel Ducret
14. Development and Validation of Multiple Reaction Monitoring (MRM) Assays for Clinical Applications
This volume presents modern and enhanced methods that detail techniques to perform proteomics analyses dedicated to biomarker discovery for human health. Chapters guide readers through pre/post analytical factors, protocols for the preparation of extracellular vesicles and exosomes, and various analytical pipelines including Data Independent Acquisition (DIA), discovery, as well as targeted and top-down proteomics analysis workflows. Bioinformatics tools and workflows to select and evaluate candidate biomarkers or combinations of biomarkers are also presented. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Proteomics for Biomarker Discovery: Methods and Protocols aims to ensure successful results in the further study of this vital field.