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This detailed volume explores robust analytical workflows for post-translational modification (PTM) analysis in both basic and translational contexts. After brief foundational reviews of the field, the book delves into detailed experimental and computational protocols for traditional bottom-up strategies and top-down proteoform analysis, as well as PTM analysis of biotherapeutic proteins, reflecting the growing emphasis on PTMs as critical quality attributes in biologic drug development and regulation. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Protein Post-Translational Modifications: Methods and Protocols serves as a foundational and forward-looking resource to support and inspire continued exploration in this rapidly evolving field.
List of contents
An Overview of Mass Spectrometry-Based Methods to Analyze Post-Translational Modifications.- Post-Translational Modifications in Protein Therapeutics: Importance and Analytical Tools.- Decoding the SUMO Proteome: A Mass Spectrometry Workflow for Site-Specific Identification.- Computational Pipelines for Protein Ubiquitylation Analysis and Prediction.- Extensive Characterization of N-Glycosylation from Major Histocompatibility Complex with Hydrophilic Interaction Chromatography and Mass Spectrometry.- Endogenous Phosphopeptidomics Analysis by Using Mesoporous IMAC Materials.- O- and N-Phosphoproteomic Analysis Using Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry.- Phosphoproteomics Protocol for the Identification of Novel Biomarkers from Extracellular Vesicles of Cancerous Cells.- Mapping Cellular Protein Lysine Methylation Using Targeted-Mass Spectrometry.- Enrichment and Metaproteomic Analysis of Lysine Acetylation in Fecal Microbiome Samples.- Generation of High-Quality Succinyl Spectral Libraries for Improved Proteome-Wide Succinylome Analysis Using Data-Independent Acquisition.- Combining SDS-PAGE with Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Top-Down Proteomics Analysis of Intact Histone Proteoforms.- PEPPI-SP3: A Gel-Based High-Resolution Sample Preparation Workflow for In-Depth Top-Down Analysis of Intact Proteoforms by Mass Spectrometry.- Profiling O-Acetylation in Sialoglycans Using MALDI-TOF and LC-MS/MS with Methylamidation and Permethylation Derivatization.- Purification of Protein-Free and Protein-Linked Poly(ADP-Ribose) from Biochemical Reactions and from Human Cells.- An NMR-Based Approach for Global Arginine Methylation Analysis.- Nanopore-Based Detection of Protein Post-Translational Modifications.- Mass Spectrometry Analysis of Biotherapeutic Monoclonal Antibody Glycosylation.- Glycoengineering of Monoclonal Antibodies with Homogeneous Glycan Isomers with Asymmetric Antennae.- Deamidation Analysis of Biotherapeutic Proteins by Capillary Electrophoresis Mass Spectrometry.
Summary
This detailed volume explores robust analytical workflows for post-translational modification (PTM) analysis in both basic and translational contexts. After brief foundational reviews of the field, the book delves into detailed experimental and computational protocols for traditional bottom-up strategies and top-down proteoform analysis, as well as PTM analysis of biotherapeutic proteins, reflecting the growing emphasis on PTMs as critical quality attributes in biologic drug development and regulation. Written for the highly successful
Methods in Molecular Biology
series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical,
Protein Post-Translational Modifications: Methods and Protocols
serves as a foundational and forward-looking resource to support and inspire continued exploration in this rapidly evolving field.
