Development of Electrospray Ionization‐Mass Spectrometry for Analysis of Water Soluble and Membrane Proteins and Educational Protocols for an Analytical Chemistry Class

Development of Electrospray Ionization‐Mass Spectrometry for Analysis of Water Soluble and Membrane Proteins and Educational Protocols for an Analytical Chemistry Class
Author: Wonhyeuk Jung
Publisher:
Total Pages: 160
Release: 2021
Genre:
ISBN:
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Native mass spectrometry (MS) is a branch of MS analysis in which the structure of the target analytes of interest are kept intact and remaining in their "native" functional structure (as much as possible). This approach was made possible by the development of electrospray ionization (ESI), a soft ionization technique that does not fragment the target analyte during the ionization process while inducing multiple charging. The multiply charged biomolecules, in turn, can be subjected to fragmentation via collisional activation with a non-reactive gas such as nitrogen. This approach of combining native MS with fragmentation-based analysis, termed native topdown MS analysis, can be applied to large biomolecules such as membrane proteins to gain structural insights. Membrane proteins present unique challenges to conventional high-resolution structural techniques due to their hydrophobic nature. However, they are responsible for various physiological phenomena and account for 60% of known druggable targets in the cell. Thus, there is a need for an approach that can overcome issues with membrane protein analysis while complementing other biophysical techniques used to probe protein structure. Here, how native top-down MS can play this role is presented. The effects of non-ionic saccharide-based detergents, a commonly used class of detergents for membrane protein solubilization, on the resulting charge states of soluble proteins is investigated to gain insights into the mechanism of ESI. The MS-fragmentation patterns from collisionally activated dissociation of membrane proteins and membrane protein-lipid complexes are compared. How new insights into the lipid binding sites can be gained by detecting lipid-bound MS-fragments is presented. The result of the study indicates that native top-down MS analysis can provide unique structural insights for membrane proteins and their non-covalent interactions. When the analytical goal is to investigate the atomic composition of the target analyte, an ionization approach in which the sample is fully atomized before MS analysis is preferred instead. Inductively coupled plasma ionization, which atomizes and ionizes the sample via a plasma, can be coupled with MS analysis (ICP-MS) to quantify heavy metal contamination in complex samples. A protocol for ICP-MS analysis of commercial fish products for mercury contamination detection developed to aid an analytical chemistry class for instruction of undergraduate chemistry students is presented.

Applied Electrospray Mass Spectrometry

Applied Electrospray Mass Spectrometry
Author: Birendra N. Pramanik
Publisher: CRC Press
Total Pages: 478
Release: 2002-02-28
Genre: Science
ISBN: 1135560439
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Discussing strategies to determine the structure and machanisms of numerous compound classics, this book covers new chemical and elctrophoretic techniques for rapid sample preconcentration and separation. It summarizes breakthroughs in the theory and instrumentation of electrospray mass spectrometry in pharmaceutical and biomedical applications, provides practical examples for the characterization of peptides, proteins, and glycoproteins, includes applications in proteomics, combinatorial chemistry, and drug characterization. Topics include chemical and electrophoretic techniques for rapid sample preconcentration and separation, screening processes for proteins from libraries of compounds, protein folding and dynamics, and more.

Electrospray and MALDI Mass Spectrometry

Electrospray and MALDI Mass Spectrometry
Author: Richard B. Cole
Publisher: John Wiley & Sons
Total Pages: 900
Release: 2011-09-26
Genre: Science
ISBN: 1118211553
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Discover how advances in mass spectrometry are fueling new discoveries across a broad range of research areas Electrospray and MALDI Mass Spectrometry brings both veteran practitioners and beginning scientists up to date with the most recent trends and findings in electrospray ionization and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. In particular, this Second Edition highlights how advances in electrospray and MALDI mass spectrometry are supporting important discoveries in new and emerging fields such as proteomics and metabolomics as well as in traditional areas of chemistry and physics research. Electrospray AND MALDI Mass Spectrometry, SECOND EDITION is divided into five parts: Part A, Fundamentals of ES, explains the fundamental phenomena underlying the electrospray process, including selectivity in ionization and inherent electrochemistry, and concludes with a chapter offering a comparative inventory of source hardware Part B, Fundamentals of MALDI, confronts ionization mechanisms, instrument development, and matrix selection, and includes a final chapter that explores the special application of MALDI to obtain two-dimensional images of spatial distributions of compounds on surfaces Part C, ES and MALDI Coupling to Mass Spectrometry Instrumentation, examines the coupling of these ionization techniques to various mass analyzers, including quadrupole ion trap, time-of-flight, Fourier transform ion cyclotron resonance, and ion mobility mass spectrometers Part D, Practical Aspects of ES and MALDI, investigates analytical issues including quantification, charge-state distributions, noncovalent interactions in solution that are preserved as gas-phase ions, and various means of ion excitation in preparation for tandem mass spectrometry, and offers a guide to the interpretation of even-electron mass spectra Part E, Biological Applications of ES and MALDI, examines the role of mass spectrometry in such areas as peptide and protein characterization, carbohydrate analysis, lipid analysis, and drug discovery Written by a team of leading experts, the book not only provides a critical review of the literature, but also presents key concepts in tutorial fashion to help readers take full advantage of the latest technological breakthroughs and applications. As a result, Electrospray and MALDI Mass Spectrometry will help researchers fully leverage the power of electrospray and MALDI mass spectrometry. The judicious compartmentalization of chapters, and the pedagogic presentation style throughout, render the book highly suitable for use as a text for graduate-level courses in advanced mass spectrometry.

Electrospray Ionization Mass Spectrometry

Electrospray Ionization Mass Spectrometry
Author: Richard B. Cole
Publisher: Wiley-Interscience
Total Pages: 610
Release: 1997-05-06
Genre: Science
ISBN:
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Comprehensive, up-to-date coverage of a revolutionary technique Electrospray ionization mass spectrometry (ESI-MS) has completely changed the way physicists, chemists, and biologists view the study of large molecules. The technique derives detailed information about molecular weights and structures from extremely small sample quantities. ESI-MS can create highly charged forms of very high molecular weight compounds, it is naturally compatible with many types of separation techniques, and it allows noncovalent interactions between molecules in solution to be preserved in the gas phase. But many researchers may not use the technique to its full potential because they are unfamiliar with the different perspectives of its underlying processes, the varied approaches to implementation, and the wide-ranging utility of the technique. In this book, Richard B. Cole and an assemblage of leading researchers present a single-volume compilation of different approaches to the understanding and exploitation of ESI-MS. This comprehensive guide: * Examines the physical and chemical aspects of the electrospray process and describes the events involved in ion formation as well as the electro-chemical phenomena that are central to charged droplet formation during the process * Explores the coupling of electrospray ionization to various mass spectrometers, including quadrupole, magnetic, time-of-flight, quadrupole ion trap, and Fourier transform ion cyclotron resonance instruments * Describes recent progress in interfacing ESI with solution-based separation techniques, including liquid chromatography and capillary electrophoresis * Charts the rapid development of ESI applications and categorizes them by compound type: peptides and proteins, nucleic acids and their constituents, carbohydrates and lipids, small molecules related to pharmacology and drug metabolism, and organometallics and inorganic compounds Electrospray Ionization Mass Spectrometry is the indispensable handbook and reference for anyone who wishes to understand, implement, or apply this technique, including researchers in chemistry, metallochemistry, biochemistry, biology, pharmacology, and physics.

Protein Analysis using Mass Spectrometry

Protein Analysis using Mass Spectrometry
Author: Mike S. Lee
Publisher: John Wiley & Sons
Total Pages: 282
Release: 2017-05-26
Genre: Science
ISBN: 1119359368
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Presents Practical Applications of Mass Spectrometry for Protein Analysis and Covers Their Impact on Accelerating Drug Discovery and Development Covers both qualitative and quantitative aspects of Mass Spectrometry protein analysis in drug discovery Principles, Instrumentation, Technologies topics include MS of peptides, proteins, and ADCs , instrumentation in protein analysis, nanospray technology in MS protein analysis, and automation in MS protein analysis Details emerging areas from drug monitoring to patient care such as Identification and validation of biomarkers for cancer, targeted MS approaches for biomarker validation, biomarker discovery, and regulatory perspectives Brings together the most current advances in the mass spectrometry technology and related method in protein analysis

Ambient Ionization Mass Spectrometry in Life Sciences

Ambient Ionization Mass Spectrometry in Life Sciences
Author: Kei Zaitsu
Publisher: Elsevier
Total Pages: 292
Release: 2019-11-06
Genre: Science
ISBN: 0128172215
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Ambient Ionization Mass Spectrometry in Life Sciences: Principles and Applications is a systematic introduction to this rapidly expanding area of study. Underlying principles of each technique are explained in detail, along with discussions on their applications across life science disciplines. Ambient ionization has recently emerged as one of the hottest and fastest growing topics in mass spectrometry, hence this book is not just for analysts and researchers who use and study mass spectrometry. This volume would be of interest to anyone who works in or studies analytical chemistry, omics sciences (including metabolomics), pharmacokinetics, forensic science or drug analysis. Covers the most up-to-date techniques, including DART, DCBI, DESI, PESI, PSI, REIMS and laser-based ambient ionization Includes easy-to-understand pros and cons of each ionization technique to aid in decision-making Provides plentiful examples of life science applications

Ambient Ionization Mass Spectrometry

Ambient Ionization Mass Spectrometry
Author: Marek Domin
Publisher: Royal Society of Chemistry
Total Pages: 528
Release: 2014-11-25
Genre: Science
ISBN: 1849739269
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Ambient ionization has emerged as one of the hottest and fastest growing topics in mass spectrometry enabling sample analysis with minimal sample preparation. Introducing the subject and explaining the basic concepts and terminology, this book will provide a comprehensive, unique treatise devoted to the subject. Written by acknowledged experts, there are full descriptions on how new ionization techniques work, with an overview of their strengths, weaknesses and applications. This title will bring the reader right up to date, with both applications and theory, and will be suitable as a tutorial text for those starting in the field from a variety of disciplines.

Fundamental Studies of Protein Ionization for Improved Analysis by Electrospray Ionization Mass Spectrometry and Related Methods

Fundamental Studies of Protein Ionization for Improved Analysis by Electrospray Ionization Mass Spectrometry and Related Methods
Author: Kevin A. Douglass
Publisher:
Total Pages: 190
Release: 2014
Genre:
ISBN:
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Mass spectrometry is an analytical technique in which a sample is converted to gas phase ions that are subsequently separated and detected. It offers great speed, selectivity, and sensitivity during analysis, characteristics which have enabled it to become a leading method for the study of proteins. The applications of MS for these biologically significant macromolecules range from accurately determining identity and sequence to shedding light on post-translational modifications and protein molecule interactions. As a first step towards analysis by MS, gas-phase protein ions must be formed. A common method for ionization is electrospray ionization, where a liquid sample including the protein is charged, nebulized, and evaporated, resulting in bare protein ions. Although ESI has been used in this way for over two decades, many aspects of the protein charging mechanism remain unclear. To address this problem, my research has focused on identifying the factors that determine the extent of protein multiple charging during ESI and improving the ionization of proteins by desorption electrospray ionization. DESI is a method similar to ESI, except that the sample is desorbed from a surface by the spray instead of being present in it from the onset. A simple model was developed that enables the accurate prediction of protein multiple charging observed during ESI-MS if the protein sequence is known. Furthermore, the enhancement of multiple charging that is observed upon the addition of certain organic reagents, a phenomenon known as supercharging, was investigated and a novel mechanism of protein supercharging was proposed. The difficulty in analyzing large proteins by DESI-MS was studied using an innovative approach where DESI was separated into its individual sub-processes and their individual contributions to the DESI process were evaluated. As a result, core limitations to the DESI-MS of large proteins were identified. The results of my cumulative research efforts should lead to the improved MS analysis of proteins by spray ionization methods, including ESI and DESI.

Biochemical and Biotechnological Applications of Electrospray Ionization Mass Spectrometry

Biochemical and Biotechnological Applications of Electrospray Ionization Mass Spectrometry
Author: A. Peter Snyder
Publisher:
Total Pages: 626
Release: 1996
Genre: Business & Economics
ISBN:
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Presents new ways of creating ions of polar and labile biopolymeric species with remarkable ease and efficiency. Introduces better characterization of proteins and other biological molecules. Presents major fundamental issues in such areas as immunology, biochemistry, and natural toxins. Demonstrates methods used to resolve the data from various biological process investigations into useful information. Provides an application-driven approach, presenting valuable insights into the experiments in the volume.