Download or read book Advancing Mass Spectrometry Methods for Quantification and Characterization of Peptides and Proteins in Complex Biological Samples written by and published by . This book was released on 2015 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein quantification using liquid chromatography mass spectrometry (LC-MS) is a critical component in the discovery and verification of protein candidate biomarkers. Label-free and label-based quantitative methods are both commonly employed, and both have advantages and disadvantages. Label-based methods are capable of analyzing relative protein abundances from multiple samples in one LC-MS experiment, thus enabling high throughput analysis. Unfortunately, the high cost of commercial labels prevents many groups from including them in their proteomics workflows. Our group previously developed a cost-effective isobaric label set, DiLeu, through dimethylation of L-leucine isotopes. The DiLeu reagent performed comparably to commercial options, but multiple improvements and variations to this reagent were needed. For example, the multiplexing capacity of the original DiLeu reagent only permits four quantitative channels, necessitating development of a new label with greater throughput. A portion of this thesis is dedicated to the design, synthesis, and characterization of a new 8-plex DiLeu reagent. This dissertation also recounts the development of isotopic DiLeu (iDiLeu), a 5-plex mass difference variant of DiLeu that allows for researchers to quantify analyte peptide concentrations from a four-point calibration curve in one LC-MS run. One glaring omission in the development of noncommercial isobaric labels is that no protein candidate biomarkers have been discovered by them. Our work rectifies this oversight by identifying several potential biomarkers using custom-synthesized 4-plex DiLeu isobaric tags from the urine of men diagnosed with lower urinary tract symptoms (LUTS). Quantitative strategies presented in this dissertation are complemented by advances in our neuropeptidomics methodologies. In one project, we study the role of protein degradation in crustacean neuropeptidomics and apply a heat stabilization strategy to mitigate the number of protein fragments that interfere with neuropeptide identification. The final project investigates an alternative method to matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) in crab neuropeptide and lipid imaging. This technique, nanostructure initiator mass spectrometry (NIMS), was unable to ionize neuropeptides in C. borealis brain tissue despite its capability to ionize lipids, showing that sometimes traditional methods like MALDI MSI remain advantageous over more recent but less vetted techniques. The future outlook for our LUTS collaboration and quantitative tag projects is positive because of the new directions still being explored. Overall, the new quantitative techniques developed in this dissertation and the advances in neuropeptide identification will benefit research groups interested in cost-effective label-based quantification methods and neuropeptide characterization in LC-MS and MSI studies.

Download or read book Mass Spectrometry of Proteins and Peptides written by John R. Chapman and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 539 pages. Available in PDF, EPUB and Kindle. Book excerpt: Little more than three years down the line and I am already writing the Preface to a second volume to follow Protein and Peptide Analysis by Mass . What has happened in between these times to make this second venture worthwhile? New types of mass spectrometric instrumentation have appeared so that new techniques have become possible and existing techniques have become much more feasible. More particularly, however, the newer ionization te- niques, introduced for the analysis of high molecular weight materials, have now been thoroughly used and studied. As a result, there has been an en- mous improvement in the associated sample handling technology so that these methods are now routinely applied to much smaller sample amounts as well as to more intractable samples. Again, this particular community of mass spectrometry users has both increased in number and diversified. And, riding this wave of acceptance, leaders in the field have set their sights on more complex problems: molecular interaction, ion structures, quantitation, and kinetics are just a few of the newer areas reported in Mass Spectrometry of Proteins and Peptides. As with the first volume, one purpose of this collection, Mass Spectr- etry of Proteins and Peptides, is to show the reader what can be done by the application of mass spectrometry, and perhaps even to encourage the reader to venture down new paths.

Download or read book Protein and Peptide Mass Spectrometry in Drug Discovery written by Michael L. Gross and published by John Wiley & Sons. This book was released on 2011-09-26 with total page 484 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book that highlights mass spectrometry and its application in characterizing proteins and peptides in drug discovery An instrumental analytical method for quantifying the mass and characterization of various samples from small molecules to large proteins, mass spectrometry (MS) has become one of the most widely used techniques for studying proteins and peptides over the last decade. Bringing together the work of experts in academia and industry, Protein and Peptide Mass Spectrometry in Drug Discovery highlights current analytical approaches, industry practices, and modern strategies for the characterization of both peptides and proteins in drug discovery. Illustrating the critical role MS technology plays in characterizing target proteins and protein products, the methods used, ion mobility, and the use of microwave radiation to speed proteolysis, the book also covers important emerging applications for neuroproteomics and antigenic peptides. Placing an emphasis on the pharmaceutical industry, the book stresses practice and applications, presenting real-world examples covering the most recent advances in mass spectrometry, and providing an invaluable resource for pharmaceutical scientists in industry and academia, analytical and bioanalytical chemists, and researchers in protein science and proteomics.

Download or read book Characterization of Protein Therapeutics using Mass Spectrometry written by Guodong Chen and published by Springer Science & Business Media. This book was released on 2014-07-08 with total page 408 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book highlights current approaches and future trends in the use of mass spectrometry to characterize protein therapies. As one of the most frequently utilized analytical techniques in pharmaceutical research and development, mass spectrometry has been widely used in the characterization of protein therapeutics due to its analytical sensitivity, selectivity, and specificity. This book begins with an overview of mass spectrometry techniques as related to the analysis of protein therapeutics, structural identification strategies, quantitative approaches, followed by studies involving characterization of process related protein drug impurities/degradants, metabolites, higher order structures of protein therapeutics. Both general practitioners in pharmaceutical research and specialists in analytical sciences will benefit from this book that details step-by-step approaches and new strategies to solve challenging problems related to protein therapeutics research and development.

Download or read book HT MS Tools for Data Quantification written by Joanne Ava and published by . This book was released on 2023-10-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Mass Spectrometry Data Analysis in Proteomics written by Rune Matthiesen and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 322 pages. Available in PDF, EPUB and Kindle. Book excerpt: This is an in-depth guide to the theory and practice of analyzing raw mass spectrometry (MS) data in proteomics. The volume outlines available bioinformatics programs, algorithms, and databases available for MS data analysis. General guidelines for data analysis using search engines such as Mascot, Xtandem, and VEMS are provided, with specific attention to identifying poor quality data and optimizing search parameters.

Download or read book Analysis of Protein Post Translational Modifications by Mass Spectrometry written by John R. Griffiths and published by John Wiley & Sons. This book was released on 2016-10-12 with total page 415 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covers all major modifications, including phosphorylation, glycosylation, acetylation, ubiquitination, sulfonation and and glycation Discussion of the chemistry behind each modification, along with key methods and references Contributions from some of the leading researchers in the field A valuable reference source for all laboratories undertaking proteomics, mass spectrometry and post-translational modification research

Download or read book Mass Spectrometry of Peptides written by Dominic M. Desiderio and published by CRC Press. This book was released on 1990-11-14 with total page 438 pages. Available in PDF, EPUB and Kindle. Book excerpt: The purpose of this book is to collect into one volume the research done on the mass spectrometry of peptides. It balances a range of topics including theory, instrumentation, analytical techniques, and biological applications. The scope of the work contains three major sections: ionization methods, instrumental developments, and analysis of peptides. It describes 252Cf plasma desorption and laser-induced multiphoton ionization methodology. This exciting resource covers many new areas, including continuous flow FAB, quantification of human neuropeptides, and peptide mapping. It also discusses Q-FTMS, cross-links, and metal ions.

Download or read book Mass Spectrometry of Proteins and Peptides written by Mary S. Lipton and published by Humana Press. This book was released on 2014-11-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: When the last edition of this book was published in 2000, the field of proteomics was in its infancy. At that time, multidimensional liquid chromatographic separations were being introduced as an alternative to traditional gel-based techniques for separating complex protein and peptide mixtures prior to mass spectrometric detection. Today, this approach – referred to as shotgun proteomics – is considered routine for lar- scale global analyses of protein mixtures. Now in its adolescence, proteomics is fundamentally transforming biological and medical research. Much of this transformation can be attributed to technological advancements, particularly in mass spectrometry. Much wider accessibility of hi- resolution and mass measurement accuracy instrumentation in recent years has ini- ated a new revolution in the field by providing more reliable data and shifting the focus from cataloging proteins to precisely quantifying changes in protein abundance over time and in response to stimuli. Advanced mass spectrometers and novel ion d- sociation schemes such as electron transfer/capture dissociation make it possible to venture boldly into the maze of protein posttranslational modifications, which are an integral component of understanding functional proteomics in the spatial and t- poral domains. Another area that has benefited from these advancements is top-down proteomics, an emerging method essential for characterizing various protein variants that has potentially high impact in biomedical research.

Download or read book Protein Analysis using Mass Spectrometry written by Mike S. Lee and published by John Wiley & Sons. This book was released on 2017-06-19 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Download or read book Selected Reaction Monitoring Mass Spectrometry SRM MS in Proteomics written by Mahmud Hossain and published by Springer Nature. This book was released on 2020-09-26 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covering a wide-ranging facet of a “gold-standard” targeted mass spectrometry (MS) method for the consistent detection and accurate quantification of preselected proteins in complex biological matrices, Selected Reaction Monitoring Mass Spectrometry (SRM-MS) in Proteomics: A Comprehensive View describes: The knowledge-based development of highly efficient SRM methodology including assay workflow, selection of proteins, peptides, transitions and its validation, and quality assessment Available bioinformatic tools – for both pre-acquisition method development and post-MS acquisition data analysis and data repositories Various relative and absolute quantification techniques SRM-MS’ widespread applications in biomarker development and in clinical studies, as well as in the analysis of various posttranslational modifications (PTMs) Current challenges and contemporary trends to overcome those difficulties In addition, it features the historical development of modern-day mass spectrometry with its vivid applications and also covers basic MS instrumentation, ionization techniques, and various proteomics approaches. Comprehensive discussion, extensive references at the end of each chapter, and the list of review articles in the bibliography offer invaluable resources for advanced readings. Researchers from the undergraduate to postgraduate level and beyond in both academic or industry settings studying and working on mass spectrometry and/or proteomics will benefit from this book.

Download or read book Mass Spectrometry Analysis for Protein Protein Interactions and Dynamics written by M. Chance and published by John Wiley & Sons. This book was released on 2008-09-22 with total page 325 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents a wide variety of mass spectrometry methods used to explore structural mechanisms, protein dynamics and interactions between proteins. Preliminary chapters cover mass spectrometry methods for examining proteins and are then followed by chapters devoted to presenting very practical, how-to methods in a detailed way. Includes footprinting and plistex specifically, setting this book apart from the competition.

Download or read book Advancing Bottom up Proteomics and Ptm Characterization Via Development of Novel Mass Spectrometry based Approaches written by Fengfei Ma (Ph.D.) and published by . This book was released on 2018 with total page 335 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry (MS) has emerged as a central technology for biomedical research largely due to its capability to provide unprecedented insights into the compositions and structures of the proteome. Thousands of proteins and peptides can be characterized in a single experiment, shedding lights on the complex biological processes and enabling discovery of novel therapeutic targets. This dissertation is devoted to the development of novel liquid chromatography (LC)-MS based techniques to characterize proteins and decipher post-translational modifications (PTMs) qualitatively and quantitatively. Specifically, novel sample preparation strategy has been developed for improved sample recovery and unbiased extraction of different categories of proteins. State-of-the-art hybrid fragmentation methods have been established for large scale differentiation of 3- and 4- hydroxyproline isomers. Furthermore, an integrated pipeline for in-depth investigation of protein glycosylation has been developed and implemented. These innovative analytical methodologies have been successfully demonstrated and applied to comprehensive characterization of biological scaffolds and breast cancer cells, which provide valuable insights into multiple research areas including tissue engineering and cancer biology.

Download or read book Quantitative Proteomics by Mass Spectrometry written by Salvatore Sechi and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 223 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume is a compendium of cutting-edge protocols for quantitative proteomics, and presents the most significant methods used in the field today. The focus on mass spectrometry (MS) is integral. Attention is given to state-of-the-art techniques for the characterization of the phosphoproteome and tandem MS for detection of inborn errors of metabolism. This volume is an indispensable resource in the search for novel biomarkers.

Download or read book Mass Spectrometry in the Biological Sciences written by A. L. Burlingame and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 569 pages. Available in PDF, EPUB and Kindle. Book excerpt: Leading practitioners describe in detail advanced methods of mass spectrometry used in structural characterization of biomacromolecules of both natural and recombinant origin. They demonstrate by example how these methodologies can solve a wide array of real-world problems in protein biochemistry, immunology, and glycobiology, as well as for human bacterial pathogens, lipids, and nucleic acids. The book offers a unique opportunity to learn these techniques that are revolutionizing the field. Its authoritative assessment in the context of how to solve important and challenging problems in bioscience and medicine ensures a competitive advantage for today's researchers.

Download or read book Introduction to Protein Mass Spectrometry written by Pradip K. Ghosh and published by Elsevier. This book was released on 2024-04-26 with total page 388 pages. Available in PDF, EPUB and Kindle. Book excerpt: Introduction to Protein Mass Spectrometry, Second Edition provides a comprehensive overview of this increasingly important, yet complex, analytical technique. This book enables readers to understand how determinations about protein identity from mass spectrometric data are made. Coverage begins with the technical basics, including preparations, instruments, and spectrometric analysis of peptides and proteins, before exploring applied use in biological applications, bioinformatics, database, and software resources. This new edition is fully updated to include the latest developments in the field and will feature new content covering recent progress in the areas where there have been the most exciting advances. These include PNNL’s multilevel-PCB-based SLIM realization, SLIM-Agilent QQQ field trials; employment of SLIM-IMS-cryo-IR combination in molecular structure determination; proximity-labelling mass spectrometry, and applications in neuroscience. Offers up-to-date, introductory information for scientists and researchers new to the field, as well as advanced insights into the critical assessment of computer-analyzed mass spectrometric results and their current limitations Provides examples of commonly used MS instruments from a range of key manufacturers/developers, including Bruker, Applied Biosystems, JEOL, Thermo Scientific/Thermo Fisher Scientific, IU, Waters and PNNL Includes biological applications and exploration of analytical tools and databases for bioinformatics Features definitions, case studies, and recent developments in protein mass spectrometry Includes sections new to this edition on SLIM (Structures for Lossless Ion Manipulation) and mass spectrometry applications in neuroscience, including synaptic biology and Alzheimer’s disease

Download or read book Advancing Intact Protein Analysis by Top down Mass Spectrometry written by Bifan Chen and published by . This book was released on 2019 with total page 215 pages. Available in PDF, EPUB and Kindle. Book excerpt: The study of proteins is critical for understanding cellular functions at the molecular level. Top-down mass spectrometry (MS) has emerged as a premier tool for global and comprehensive analysis of proteoforms. The top-down approach retains intact mass information, providing a "bird's-eye" view of the proteome and allowing for identification of novel proteoforms, in-depth sequence characterization, and quantification of disease associated post-translational modifications (PTMs). However, many technical challenges still exist. The research described here involves analytical development in top-down MS, particularly in the areas of enrichment, separation, and characterization of samples ranging from standard proteins and complex lysates, to large therapeutic biomolecules. Chapter 1 provides an introduction and review of recent advances in different aspects of top-down proteomics. Chapters 2 and 3 are related to the study of intact phosphoproteins. Specifically, chapter 2 describes the use of functionalized nanoparticles for enrichment and the subsequent coupling of online liquid chromatography (LC)-MS for characterizing endogenous phosphoproteins from complex cell lysates. Chapter 3 investigates how phosphorylation moieties might influence the efficiency of electron capture dissociation (ECD). Chapters 4 and 5 focus on the development of hydrophobic interaction chromatography (HIC) that could be coupled online directly with MS and its applications to therapeutic molecules (monoclonal antibodies). Chapter 6 describes a middle-down approach to obtain multi-attribute of both cysteine and lysine conjugated antibody-drug conjugates, which overcomes some current challenges using HIC-MS and the top-down approach. Overall, these analytical developments expand the toolbox of the top-down approach and generally facilitate the analysis of intact proteins.