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Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics A powerful and essential resource for researchers with an interest in CE-MS In Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics: Principles and Applications, a team of distinguished researchers delivers a comprehensive overview of bioanalytical capillary electrophoresis coupled to mass spectrometry (CE-MS). The book explains foundational principles, technology as well the strategies and techniques used in data analysis for metabolic and proteomic studies. It also provides a global overview of recent developments and advances for improving CE-MS sensitivity and reproducibility. An essential handbook for everyone performing metabolomic and proteomic analysis, the information provided here will assist researchers in tapping into the full potential of this technique to answer biological and clinical questions. Readers will also find: A thorough introduction to the principles of capillary electrophoresis, including its fundamentals, CE separation modes, capillary coatings, and the fundamentals of mass spectrometry In-depth examinations of technological developments in capillary electrophoresis, including sample preparation, online preconcentration, detection sensitivity, and metabolic coverage Comprehensive discussions of metabolomic studies, including their biomedical and clinical applications Recent advances in proteomics, including top-down and bottom-up approaches Perfect for analytical and clinical chemists, Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics: Principles and Applications will also earn a place in the libraries of biochemists, molecular biologists, and other molecular life scientists.
Diese Monographie bietet einen vollständigen Überblick über die Prinzipien und Anwendungen der Kapillarelektrophorese (CE) und der Massenspektrometrie (MS) und legt den Nachdruck insbesondere auf Kopplungsschnittstellen. Ausführlich erläutert werden auch alle relevanten Substanzklassen. Ein einzigartiges Wissenskompendium für alle, die sich CE-MS beschäftigen!
Capillary electrophoresis–mass spectrometry (CE-MS) has become a very useful analytical technique for the profiling of highly polar and charged metabolites in biological samples. In this book, the unique features of CE-MS for metabolomics studies are highlighted including CE separation modes, capillary coatings and practical aspects of CE-MS coupling alongside a comprehensive overview of recent technological developments and applications. CE-MS can be considered a relatively new technique in the field of metabolomics and it is therefore important to inform the scientific community about the possibilities of advanced CE-MS approaches for metabolomics studies. This book outlines the potential of this technique for researchers working in metabolomics, bioanalytics and biomarker analysis.
This monograph offers the reader a complete overview on both principles and applications of CE-MS. Starting with an introductory chapter on detection in CE, also related and more specialized techniques such as electrophoretic and chromatographic preconcentration are discussed. A special emphasis is put on CE-MS interfaces, which are described in detail. In a separate chapter, attention is paid to sheath-liquid interfacing. The developments and possibilities of microchip CE-MS are also described. Applications to all relevant areas are discussed in distinct chapters, each written by experts in the respective fields. Besides applications in pharmaceutical analysis and bioanalysis, recent implementations in food science, forensic analysis, analysis of intact proteins, metabolomics and proteomics are highlighted. MS is a perfectly appropriate detection system for CE, as efficient separation is coupled to sensitive and selection detection. Moreover, MS can provide structure information on the separated compounds. CE-MS has now been developed into a strong hyphenated system complementary to LC-MS. This monograph is an unique source of knowledge for everyone dealing with and interested in CE-MS.
Capillary electrophoresis (CE) has become an established method with widespread recognition as an analytical technique of choice in numerous analytical laboratories, including industrial and academic sectors. Pharmaceutical and biochemical research and quality control are the most important CE applications. This book provides a comparative assessment of related techniques on mode selection, method development, detection, and quantitative analysis and estimation of pharmacokinetic parameters and broadens the understanding of modern CE applications, developments, and prospects. It introduces the fundamentals of CE and clearly outlines the procedures used to mitigate several barriers, such as detection limits, signal detection, changing capillary environment, resolution separation of analytes, and hyphenation of mass spectrometry with CE, for a range of analytical problems. Each chapter outlines a specific electrophoretic variant with detailed instructions and some standard operating procedures. In this respect, the book meets its desired goal of rendering assistance to lovers of electrophoresis.
This book covers the latest developments in capillary electrophoresis-mass spectrometry for the analysis of therapeutic proteins. The application of capillary electrophoresis-mass spectrometry (CE-MS) coupling technology in the analysis of recombinant therapeutic proteins is detailed thoroughly. Specific topics include recent developments in coupling capillary electrophoresis with mass spectrometry for the quality control of monoclonal antibody therapeutics, top-down analysis of monoclonal antibody using the CE-MS platform, and detection of host cell protein impurities. Comprehensive characterization of antibody-drug conjugates (ADCs) by coupling capillary electrophoresis with mass spectrometry is also covered. This is an ideal book for scientists in the life science and biopharmaceutical industry who are working on characterizing the PTMs of monoclonal antibodies, as well as graduate students and researchers in the separation science and biological mass spectrometry fields.
Due to the high sensitivity, selectivity, and the possibility for detailed molecular characterisation, mass spectrometry (MS) is the analytical method of choice for top-down protein biomarker discovery. Due to the low sample consumption, high separation efficiency, and the unique and complementary selectivity, capillary zone electrophoresis (CZE)-MS represents an interesting alternative to the traditionally used liquid chromatography (LC)-MS platforms. In this thesis, instrumental and methodological concepts were developed to increase the potential of CZE-MS for intact proteins. The first part of the thesis describes the development of a nanoflow sheath liquid interface for the efficient coupling of CZE and MS. The interface was developed with a focus on fast setup, easy handling and analytical robustness and has been used for most applications in this thesis. Furthermore, a CZE-MS screening platform for the identification and characterisation of known and unknown Hb variants from DBS samples was developed. The application of SMIL coatings enables efficient separation of closely-related proteoforms and even positional isomers of glycated Hb on the intact level. In the last part of the thesis, nanoLC and CZE-MS were coupled in a heart-cut approach using a polymer nanoliter valve. The platform was used for the glycosylation profiling of heterogeneous alpha-1 acid glycoprotein (AGP). This approach enables the assignment of notably more glycoforms from a lower concentrated AGP sample, compared to CZE-MS alone. In a proof-of-concept study, the platform was further extended to operate in the selective comprehensive mode. With a single injection, 19% more glycoforms were assigned compared to the heart-cut approach with 3 injections. The here presented instrumental and methodological concepts show the great potential of CZE-MS in the context of clinical protein analysis. Especially the combination of LC and CZE in multidimensional separation platforms shows great potential.
This book presents a selection of current capillary electrophoresis methods used to separate representative types of molecules and particles and in combination with different detection techniques. It includes practical details which are hard to find elsewhere. The volume is intended for beginners in the field and provides an overview of the technique and a starting point for the exploration of the defined literature on different application topics.