Method Development For The Analysis Of Proteins And Small Molecules By Ce Lif Ce Ms And Hplc Elsd

Download Method Development For The Analysis Of Proteins And Small Molecules By Ce Lif Ce Ms And Hplc Elsd full books in PDF, epub, and Kindle. Read online Method Development For The Analysis Of Proteins And Small Molecules By Ce Lif Ce Ms And Hplc Elsd ebook anywhere anytime directly on your device. Fast Download speed and no annoying ads. We cannot guarantee that every ebooks is available!

Method Development for the Analysis of Proteins and Small Molecules by CE-LIF, CE-MS, and HPLC-ELSD

Author : Anthony Richard Gerardi
Publisher :
ISBN 13 :
Total Pages : 157 pages
Book Rating : 4.:/5 (812 download)

DOWNLOAD NOW!

Book Synopsis Method Development for the Analysis of Proteins and Small Molecules by CE-LIF, CE-MS, and HPLC-ELSD by : Anthony Richard Gerardi

Download or read book Method Development for the Analysis of Proteins and Small Molecules by CE-LIF, CE-MS, and HPLC-ELSD written by Anthony Richard Gerardi and published by . This book was released on 2011 with total page 157 pages. Available in PDF, EPUB and Kindle. Book excerpt: Analytical separations in conjunction with powerful detection methods make possible the quantitation of large and small molecules important in environmental science, food science, pharmaceutical, and medical matrices. Many of these separations utilize instrumental methods such as capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) to alleviate the complexity of these various matrices. Fluorescence, laser-induced fluorescence (LIF), evaporative light scattering detection (ELSD), and mass spectrometry (MS) are detection methods commonly employed in conjunction with these separation strategies. Three distinct projects to develop and exploit such analytical technologies are described. First, few biological molecules are natively fluorescent, so they often must be labeled with a fluorescent tag in order to be detected via LIF. Furthermore, protein separations using CE-LIF may be complicated by protein interactions with the negatively charged surface of the inner capillary wall, arising from the ionization of surface silanol groups. Presented here is a method for quantitation of protein-dye complexes using a self assembled, surfactant-coated capillary and on-column protein labeling with a non-covalent squarylium dye, Red-1c. Second, seasonal sugar levels are relevant to the relationship between winter anthocyanin production and drought stress in angiosperm evergreen species, and so the development of a method for rapid analysis of sugars in seasonal tree leaf tissue is needed. An HPLC-ELSD method was developed for the quantitation of three sugars, glucose, fructose, and sucrose. The method was subjected to standard addition validation and was shown to provide precise and accurate measurement of all three analytes in real samples. Third, in addition to their many uses as solvents, ionic liquids (IL) have been utilized as highly effective substrates for complexation and subsequent quantitation of anions by electrospray ionization mass spectrometry (ESI-MS) and high performance liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). The present work provides an alternative method utilizing capillary electrophoresis coupled with electrospray ionization mass spectrometry (CE-ESI-MS). This method offers advantages over LC-MS in that CE, in general, provides shorter separation times, requires less reagent and solvent, and needs less sample volume. Mono- and divalent anionic analytes, were complexed with di- and trivalent cationic liquids. The complexation reaction was undertaken in a variety of modes, including pre-column, on-column and post-column, and the resulting complexes, which were positively charged and of higher mass than the native analytes, were subsequently analyzed by MS. This method allowed for greater sensitivity than could be achieved by direct detection.

Handbook of LC-MS Bioanalysis

Author : Wenkui Li
Publisher : John Wiley & Sons
ISBN 13 : 111867135X
Total Pages : 709 pages
Book Rating : 4.1/5 (186 download)

DOWNLOAD NOW!

Book Synopsis Handbook of LC-MS Bioanalysis by : Wenkui Li

Download or read book Handbook of LC-MS Bioanalysis written by Wenkui Li and published by John Wiley & Sons. This book was released on 2013-09-03 with total page 709 pages. Available in PDF, EPUB and Kindle. Book excerpt: Consolidates the information LC-MS bioanalytical scientists need to analyze small molecules and macromolecules The field of bioanalysis has advanced rapidly, propelled by new approaches for developing bioanalytical methods, new liquid chromatographic (LC) techniques, and new mass spectrometric (MS) instruments. Moreover, there are a host of guidelines and regulations designed to ensure the quality of bioanalytical results. Presenting the best practices, experimental protocols, and the latest understanding of regulations, this book offers a comprehensive review of LC-MS bioanalysis of small molecules and macromolecules. It not only addresses the needs of bioanalytical scientists working on routine projects, but also explores advanced and emerging technologies such as high-resolution mass spectrometry and dried blood spot microsampling. Handbook of LC-MS Bioanalysis features contributions from an international team of leading bioanalytical scientists. Their contributions reflect a review of the latest findings, practices, and regulations as well as their own firsthand analytical laboratory experience. The book thoroughly examines: Fundamentals of LC-MS bioanalysis in drug discovery, drug development, and therapeutic drug monitoring The current understanding of regulations governing LC-MS bioanalysis Best practices and detailed technical instructions for LC-MS bioanalysis method development, validation, and stability assessment of analyte(s) of interest Experimental guidelines and protocols for quantitative LC-MS bioanalysis of challenging molecules, including pro-drugs, acyl glucuronides, N-oxides, reactive compounds, and photosensitive and autooxidative compounds With its focus on current bioanalytical practice, Handbook of LC-MS Bioanalysis enables bioanalytical scientists to develop and validate robust LC-MS assay methods, all in compliance with current regulations and standards.

Capillary Electrophoresis-mass Spectrometry Methods for Advancements in Proteomics

Author : Selynda Ana Garza
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (166 download)

DOWNLOAD NOW!

Book Synopsis Capillary Electrophoresis-mass Spectrometry Methods for Advancements in Proteomics by : Selynda Ana Garza

Download or read book Capillary Electrophoresis-mass Spectrometry Methods for Advancements in Proteomics written by Selynda Ana Garza and published by . This book was released on 2007 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Sample Preparation in LC-MS Bioanalysis

Author : Wenkui Li
Publisher : John Wiley & Sons
ISBN 13 : 111927429X
Total Pages : 384 pages
Book Rating : 4.1/5 (192 download)

DOWNLOAD NOW!

Book Synopsis Sample Preparation in LC-MS Bioanalysis by : Wenkui Li

Download or read book Sample Preparation in LC-MS Bioanalysis written by Wenkui Li and published by John Wiley & Sons. This book was released on 2019-03-12 with total page 384 pages. Available in PDF, EPUB and Kindle. Book excerpt: Revised and Expanded Handbook Provides Comprehensive Introduction and Complete Instruction for Sample Preparation in Vital Category of Bioanalysis Following in the footsteps of the previously published Handbook of LC-MS Bioanalysis, this book is a thorough and timely guide to all important sample preparation techniques used for quantitative Liquid Chromatography–Mass Spectrometry (LC-MS) bioanalysis of small and large molecules. LC-MS bioanalysis is a key element of pharmaceutical research and development, post-approval therapeutic drug monitoring, and many other studies used in human healthcare. While advances are continually being made in key aspects of LC-MS bioanalysis such as sensitivity and throughput, the value of research/study mentioned above is still heavily dependent on the availability of high-quality data, for which sample preparation plays the critical role. Thus, this text provides researchers in industry, academia, and regulatory agencies with detailed sample preparation techniques and step-by-step protocols on proper extraction of various analyte(s) of interest from biological samples for LC-MS quantification, in accordance with current health authority regulations and industry best practices. The three sections of the book with a total of 26 chapters cover topics that include: Current basic sample preparation techniques (e.g., protein precipitation, liquid-liquid extraction, solid-phase extraction, salting-out assisted liquid-liquid extraction, ultracentrifugation and ultrafiltration, microsampling, sample extraction via electromembranes) Sample preparation techniques for uncommon biological matrices (e.g., tissues, hair, skin, nails, bones, mononuclear cells, cerebrospinal fluid, aqueous humor) Crucial aspects of LC-MS bioanalytical method development (e.g., pre-analytical considerations, derivation strategies, stability, non-specific binding) in addition to sample preparation techniques for challenging molecules (e.g., lipids, peptides, proteins, oligonucleotides, antibody-drug conjugates) Sample Preparation in LC-MS Bioanalysis will prove a practical and highly valuable addition to the reference shelves of scientists and related professionals in a variety of fields, including pharmaceutical and biomedical research, mass spectrometry, and analytical chemistry, as well as practitioners in clinical pharmacology, toxicology, and therapeutic drug monitoring.

Protein and Peptide Analysis by LC-MS

Author : Thomas Letzel
Publisher : Royal Society of Chemistry
ISBN 13 : 1849733147
Total Pages : 195 pages
Book Rating : 4.8/5 (497 download)

DOWNLOAD NOW!

Book Synopsis Protein and Peptide Analysis by LC-MS by : Thomas Letzel

Download or read book Protein and Peptide Analysis by LC-MS written by Thomas Letzel and published by Royal Society of Chemistry. This book was released on 2011-07-22 with total page 195 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is the first example in presenting LC-MS strategies for the analysis of peptides and proteins with detailed information and hints about the needs and problems described from experts on-the-job. The best advantage is -for sure- the practical insight of experienced analysts into their novel protein analysis techniques. Readers starting in 'Proteomics' should be able to repeat each experiment with own equipment and own protein samples, like clean-up, direct protein analysis, after (online) digest, with modifications and others. Furthermore, the reader will learn more about strategies in protein analysis, like quantitative analysis, industrial standards, functional analysis and more.

Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection

Author : Michael D. Harvey
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (11 download)

DOWNLOAD NOW!

Book Synopsis Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection by : Michael D. Harvey

Download or read book Nanoscale Protein Analysis Utilizing Capillary Electrophoresis and Laser-induced Fluorescence Detection written by Michael D. Harvey and published by . This book was released on 2001 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The trend towards high throughput applications and miniaturization necessitates approaches capable of microlitre volume sampling and low protein concentration detection. Furthermore, one of the major trends in high throughput screening is the growing replacement of technologies that depend on radioactivity to generate a signal with those that rely on fluorescence. This trend towards non-radioactive detection in general can be understood by some of the advantages inherent to these methods over radioactive modes. These include a significant reduction in safety concerns leading to a relaxation of strict laboratory procedures, elimination of expensive waste disposal, extended shelf-life of labeled reagents, and the possibility of acquiring multiplexed data through the spectral isolation of different wavelength signals. A variety of capillary electrophoretic (CE) approaches utilizing laser-induced fluorescence (LIF) have thus been developed, providing researchers with valuable tools in protein analysis. Various covalent and non-covalent fluorescent derivatization approaches have been investigated, with emphasis on biochemical and/or clinical applications. The non-covalent dye, NanoOrange, is used as a clinical diagnostic tool for early disease diagnosis, quantitating nanomolar concentrations of human serum albumin in solution, and obtaining fluorescence-based biofluid profiles. An alternate non-covalent labeling approach utilizing the fluorescent probe, Sypro Red, and capillary gel electrophoresis allows for rapid, sensitive analysis of protein sample purity as well as molecular weight determination. These two non-covalent approaches are complemented by the development of a fluorescent Insulin-Like Growth Factor-I (IGF-I) analog for use in bioanalytical applications. Specific derivatization reaction conditions were developed to selectively label the N-terminus of the analog hence preserve biological activity. High-performance liquid chromatography and electrospray mass spectrometry were used to confirm the extent of labeling and modification site. Antibody recognition of this fluorescent analog was evaluated using CE-LIF, illustrating the clinical utility of this diagnostic reagent. In addition to the above CE-LIF approaches, a fourth capillary electrophoretic tool is provided for the clinical chemist. Rapid analysis of biofluids is of significant importance in early disease diagnosis. As such, an extensive CE-based analysis of human seminal plasma is presented. Separation conditions, sample stability, and protein/non-protein zone identification issues are addressed. This study and the CE-LIF methodologies discussed above represent original approaches to nanoscale protein analysis.

Capillary Electrophoresis–Mass Spectrometry for Metabolomics

Author : Rawi Ramautar
Publisher : Royal Society of Chemistry
ISBN 13 : 1788014847
Total Pages : 300 pages
Book Rating : 4.7/5 (88 download)

DOWNLOAD NOW!

Book Synopsis Capillary Electrophoresis–Mass Spectrometry for Metabolomics by : Rawi Ramautar

Download or read book Capillary Electrophoresis–Mass Spectrometry for Metabolomics written by Rawi Ramautar and published by Royal Society of Chemistry. This book was released on 2018-07-12 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Capillary Electrophoresis Mass Spectrometry for Proteomics and Metabolomics

Author : Rawi Ramautar
Publisher : John Wiley & Sons
ISBN 13 : 3527349219
Total Pages : 404 pages
Book Rating : 4.5/5 (273 download)

DOWNLOAD NOW!

Book Synopsis Capillary Electrophoresis Mass Spectrometry for Proteomics and Metabolomics by : Rawi Ramautar

Download or read book Capillary Electrophoresis Mass Spectrometry for Proteomics and Metabolomics written by Rawi Ramautar and published by John Wiley & Sons. This book was released on 2022-10-11 with total page 404 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Advanced Methods in Protein Microsequence Analysis

Author : Brigitte Wittmann-Liebold
Publisher : Springer Science & Business Media
ISBN 13 : 3642715346
Total Pages : 427 pages
Book Rating : 4.6/5 (427 download)

DOWNLOAD NOW!

Book Synopsis Advanced Methods in Protein Microsequence Analysis by : Brigitte Wittmann-Liebold

Download or read book Advanced Methods in Protein Microsequence Analysis written by Brigitte Wittmann-Liebold and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 427 pages. Available in PDF, EPUB and Kindle. Book excerpt: Much of the recent spectacular progress in the biological sciences can be at tributed ot the ability to isolate, analyze, and structurally characterize proteins and peptides which are present in cells and cellular organelles in only very small amounts. Recent advances in protein chemistry and in particular the application of new micromethods have led to fruitful advances in the understanding of basic cellular processes. Areas where protein-chemical studies have resulted in interest ing discoveries include the peptide hormones and their release factors, growth factors and oncogenes, bioenergetics, proton pumps and ion pumps and chan nels, topogenesis and protein secretion, molecular virology and immunology, membrane protein analysis, and receptor research. In fact, the key methods are now on hand to unravel many of the major outstanding problems of molecular biology and in particular questions of fundamental interest which relate to devel opmental biology and specificity in cell-cell interaction. In this volume we have assembled descriptions of procedures which have re cently been shown to be efficaceous for the isolation, purification, and chemical characterization of proteins and peptides that are only available in minute amounts. Emphasis is placed on well-established micromethods which have been tested and found useful in many laboratories by experienced investigators. The chapters are written by specialists, and describe a range of sensitive techniques which can be used by researchers working in laboratories with only modest resources and equipment.

Development of On-line Capillary Electrophoresis and Monolithic Capillary HPLC Interfaced with Mass Spectrometry for Analysis of Human Cancer Proteomes

Download Development of On-line Capillary Electrophoresis and Monolithic Capillary HPLC Interfaced with Mass Spectrometry for Analysis of Human Cancer Proteomes PDF Online Free

Author : Chul Seung Yoo
Publisher :
ISBN 13 :
Total Pages : 410 pages
Book Rating : 4.3/5 (91 download)

DOWNLOAD NOW!

Book Synopsis Development of On-line Capillary Electrophoresis and Monolithic Capillary HPLC Interfaced with Mass Spectrometry for Analysis of Human Cancer Proteomes by : Chul Seung Yoo

Download or read book Development of On-line Capillary Electrophoresis and Monolithic Capillary HPLC Interfaced with Mass Spectrometry for Analysis of Human Cancer Proteomes written by Chul Seung Yoo and published by . This book was released on 2006 with total page 410 pages. Available in PDF, EPUB and Kindle. Book excerpt:

LC-MS/MS in Proteomics

Author : Pedro R. Cutillas
Publisher : Humana
ISBN 13 : 9781493957446
Total Pages : 357 pages
Book Rating : 4.9/5 (574 download)

DOWNLOAD NOW!

Book Synopsis LC-MS/MS in Proteomics by : Pedro R. Cutillas

Download or read book LC-MS/MS in Proteomics written by Pedro R. Cutillas and published by Humana. This book was released on 2016-08-23 with total page 357 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the development of new quantitative strategies and powerful bioinformatics tools to cope with the analysis of the large amounts of data generated in proteomics experiments, liquid chromatography with tandem mass spectrometry (LC-MS/MS) is making possible the analysis of proteins on a global scale, meaning that proteomics can now start competing with cDNA microarrays for the analysis of whole genomes. In LC-MS/MS in Proteomics: Methods and Applications, experts in the field provide protocols and up-to-date reviews of the applications of LC-MS/MS, with a particular focus on MS-based methods of protein and peptide quantification and the analysis of post-translational modifications. Beginning with overviews of the use of LC-M/MS in protein analysis, the book continues with topics such as protocols for the analysis of post-translational modifications, with particular focus on phosphorylation and glycosylation, popular techniques for quantitative proteomics, such as multiple reaction monitoring, metabolic labelling, and chemical tagging, biomarker discovery in biological fluids, as well as novel applications of LC-MS/MS. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective subjects, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, LC-MS/MS in Proteomics: Methods and Applications presents the techniques and concepts necessary in order to aid proteomic practitioners in the application of LC-MS/MS to essentially any biological problem.

Protein Analysis using Mass Spectrometry

Author : Mike S. Lee
Publisher : John Wiley & Sons
ISBN 13 : 1119359368
Total Pages : 282 pages
Book Rating : 4.1/5 (193 download)

DOWNLOAD NOW!

Book Synopsis Protein Analysis using Mass Spectrometry by : Mike S. Lee

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 2017-05-26 with total page 282 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

Development of Capillary Electrophoresis and Mass Spectrometry Methods in Protein Analysis

Author : Lin Liu
Publisher :
ISBN 13 :
Total Pages : 242 pages
Book Rating : 4.3/5 (91 download)

DOWNLOAD NOW!

Book Synopsis Development of Capillary Electrophoresis and Mass Spectrometry Methods in Protein Analysis by : Lin Liu

Download or read book Development of Capillary Electrophoresis and Mass Spectrometry Methods in Protein Analysis written by Lin Liu and published by . This book was released on 1999 with total page 242 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Methods for Protein Analysis

Author : Robert A. Copeland
Publisher : Springer Science & Business Media
ISBN 13 : 1475715056
Total Pages : 238 pages
Book Rating : 4.4/5 (757 download)

DOWNLOAD NOW!

Book Synopsis Methods for Protein Analysis by : Robert A. Copeland

Download or read book Methods for Protein Analysis written by Robert A. Copeland and published by Springer Science & Business Media. This book was released on 2013-11-11 with total page 238 pages. Available in PDF, EPUB and Kindle. Book excerpt: As protein science continues to become an increasingly important aspect of academic and commercial sciences and technology, the need has arisen for a ready source of laboratory protocols for the analysis and evaluation of these biological polymers. Methods for Protein Analysis presents the methods most relevant to the generalist bench scientist working with proteins. A concise yet thorough summary, it covers laboratory methods that can be reasonably performed in a standard protein laboratory, without specialized equipment or expertise. Taking a how to approach, this book examines the techniques used to answer common protein analytical questions and describes methods useful in daily laboratory work. Methods for Protein Analysis is the ideal reference for protein laboratories in academic, government and industrial settings. It is an essential benchtop manual for first-year graduate students beginning their laboratory experience as well as for chemists, biochemists, and molecular biologists in the pharmaceutical, biotechnological, food and specialty chemical industries, and for analysts concerned with the purity and structural integrity of protein. Featuring illustrations and a convenient spiral binding, this guide offers a glossary of common abbreviations and a list of suppliers for protein science.

Liquid Chromatography and Mass Spectrometry Based Analytical Method Development Towards Fast and Sensitive Analysis

Author : Juan Bian
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (118 download)

DOWNLOAD NOW!

Book Synopsis Liquid Chromatography and Mass Spectrometry Based Analytical Method Development Towards Fast and Sensitive Analysis by : Juan Bian

Download or read book Liquid Chromatography and Mass Spectrometry Based Analytical Method Development Towards Fast and Sensitive Analysis written by Juan Bian and published by . This book was released on 2019 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Liquid chromatography (LC) and mass spectrometry (MS) is a valuable tool in pharmaceutical analysis and is widely applied to the analysis of many real-world samples. Recent achievements in material and instrumentation have allowed LC and MS to play a critical role in the bioanalytical application. However, when dealing with a sample with high complexity, such as metabolites or proteins from biological samples, current LC and MS-based methods may not provide sufficient analytical results. As a consequence, this dissertation focuses on new LC and MS-based method development to address the challenges in small molecules and macromolecules analysis.

Protein Analysis and Purification

Author : I.M. Rosenberg
Publisher : Springer Science & Business Media
ISBN 13 : 1475711085
Total Pages : 453 pages
Book Rating : 4.4/5 (757 download)

DOWNLOAD NOW!

Book Synopsis Protein Analysis and Purification by : I.M. Rosenberg

Download or read book Protein Analysis and Purification written by I.M. Rosenberg and published by Springer Science & Business Media. This book was released on 2013-03-14 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

Mass Spectrometry-based Strategies for Protein Characterization

Author : Ke Li (Chemist)
Publisher :
ISBN 13 :
Total Pages : 189 pages
Book Rating : 4.:/5 (19 download)

DOWNLOAD NOW!

Book Synopsis Mass Spectrometry-based Strategies for Protein Characterization by : Ke Li (Chemist)

Download or read book Mass Spectrometry-based Strategies for Protein Characterization written by Ke Li (Chemist) and published by . This book was released on 2018 with total page 189 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry (MS)-based protein footprinting characterizes protein structure and protein-ligand interactions by interrogating protein solvent-accessible surfaces by using chemical reagents as probes. The method is highly applicable to protein or protein-ligand complexes that are difficult to study by conventional means such as X-ray crystallography and nuclear magnetic resonance. In this dissertation, we describe the development and application of MS-based protein footprinting from three perspectives, including I) protein aggregation and amyloid formation (Chapter 2-3), II) protein-ligand interactions (Chapter 4-5), and III) in-cellulo structures and dynamic motion of membrane proteins (Chapter 6). Fast Photochemical Oxidation of Proteins (FPOP) is the main methodology implemented in the work presented in this dissertation. Chapter 1 provides an overview of FPOP and discusses its fundamentals as well as its important applications in both academic research and biotechnology drug development. In Part I, Chapter 2 covers the early method development of FPOP for monitoring amyloid beta (A[beta]) aggregation. In this work, we demonstrated the high sensitivity and spatial resolution of the method in probing the solvent accessibility of A[beta] at global, sub-regional, and some amino-acid residue levels as a function of its aggregation, and revealed A[beta] species at various oligomeric states identified by their characteristic modification levels. In Chapter 3, we extended the application of the platform to assess the effect of a putative polyphenol inhibitor on amyloid formation and to provide insights into the mechanism of action of the inhibitor in remodeling A[beta] aggregation pathways. In Part II, we evaluated different protein footprinting techniques, including FPOP, hydrogen-deuterium exchange (HDX), and carboxyl group footprinting, for probing protein-ligand (drug candidates) interaction in the context of a therapeutic development. Chapter 4 focused on protein-protein interaction by investigating the epitope of IL-6 receptor for two adnectins that have similar apparent biophysical properties. In Chapter 5, we probed the hydrophobic binding cavity of bromodomain protein for a small molecule inhibitor. This study serves as an example of interrogating protein-small molecule interactions. The two studies in Part II demonstrate the unique capabilities and limitations of protein footprinting methods in protein structural characterization. In Part III, we pushed the boundary of MS-based protein footprinting by applying the method to footprint live cells and investigate the dynamic structures/motion of membrane-transport proteins in their native cellular environment. We employed protein engineering, suspension cell expression and isotopic-encoded carboxyl group footprinting to identify salt bridges in two proteins, GLUT1 and GLUT5, that control their alternating access motions for substrate translocation. With functional analysis and mutagenesis, live-cell footprinting provides new insights into the transport mechanism of proteins in the major facilitator superfamily. The five studies in the dissertation demonstrate the powerful capability of MS-based protein footprinting in protein structural biology and biophysics research. The method also holds great potential in studying more complicated biological systems and solving demanding problems related to protein structure and properties.