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…mehr
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 toensure successful results in the further study of this vital field.
Pre- and Post-Analytical Factors in Biomarker Discovery.- Pre-Fractionation of Non-Circulating Biological Fluids to Improve Discovery of Clinically Relevant Protein Biomarkers.- Serum Exosome Isolation by Size-Exclusion Chromatography for the Discovery and Validation of Preeclampsia-Associated Biomarkers.- Protein Biomarker Discovery Using Human Blood Plasma Microparticles.- A Standardized and Reproducible Proteomics Protocol for Bottom-Up Quantitative Analysis of Protein Samples Using SP3 and Mass Spectrometry.- Analyzing Cerebrospinal Fluid Proteomes to Characterize Central Nervous System Disorders: A Highly Automated Mass Spectrometry-Based Pipeline for Biomarker Discovery.- Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies.- Tube-Gel: A Fast and Effective Sample Preparation Method for High-Throughput Quantitative Proteomics.- Protein Biomarker Discovery in Non-Depleted Serum by Spectral Library-Based Data-Independent Acquisition Mass Spectrometry.- Discovering Protein Biomarkers from Clinical Peripheral Blood Mononuclear Cells Using Data-Independent Acquisition Mass Spectrometry.- Intact Protein Analysis by LC-MS for Characterizing Biomarkers in Cerebrospinal Fluid.- Detection of Proteoforms Using Top-Down Mass Spectrometry and Diagnostic Ions.- Development of a Highly Multiplexed SRM Assay for Biomarker Discovery in Formalin-Fixed Paraffin-Embedded Tissues.- Development and Validation of Multiple Reaction Monitoring (MRM) Assays for Clinical Applications.- Protein-Level Statistical Analysis of Quantitative Label-Free Proteomics Data with ProStaR.- Computation and Selection of Optimal Biomarker Combinations by Integrative ROC Analysis using CombiROC.- PanelomiX for the Combination of Biomarkers.- Designing an In Silico Strategy to Select Tissue-Leakage Biomarkers Using the Galaxy Framework.
Pre- and Post-Analytical Factors in Biomarker Discovery.- Pre-Fractionation of Non-Circulating Biological Fluids to Improve Discovery of Clinically Relevant Protein Biomarkers.- Serum Exosome Isolation by Size-Exclusion Chromatography for the Discovery and Validation of Preeclampsia-Associated Biomarkers.- Protein Biomarker Discovery Using Human Blood Plasma Microparticles.- A Standardized and Reproducible Proteomics Protocol for Bottom-Up Quantitative Analysis of Protein Samples Using SP3 and Mass Spectrometry.- Analyzing Cerebrospinal Fluid Proteomes to Characterize Central Nervous System Disorders: A Highly Automated Mass Spectrometry-Based Pipeline for Biomarker Discovery.- Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies.- Tube-Gel: A Fast and Effective Sample Preparation Method for High-Throughput Quantitative Proteomics.- Protein Biomarker Discovery in Non-Depleted Serum by Spectral Library-Based Data-Independent Acquisition Mass Spectrometry.- Discovering Protein Biomarkers from Clinical Peripheral Blood Mononuclear Cells Using Data-Independent Acquisition Mass Spectrometry.- Intact Protein Analysis by LC-MS for Characterizing Biomarkers in Cerebrospinal Fluid.- Detection of Proteoforms Using Top-Down Mass Spectrometry and Diagnostic Ions.- Development of a Highly Multiplexed SRM Assay for Biomarker Discovery in Formalin-Fixed Paraffin-Embedded Tissues.- Development and Validation of Multiple Reaction Monitoring (MRM) Assays for Clinical Applications.- Protein-Level Statistical Analysis of Quantitative Label-Free Proteomics Data with ProStaR.- Computation and Selection of Optimal Biomarker Combinations by Integrative ROC Analysis using CombiROC.- PanelomiX for the Combination of Biomarkers.- Designing an In Silico Strategy to Select Tissue-Leakage Biomarkers Using the Galaxy Framework.
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