Download or read book The Use of 2D LC MS MS in Disease Characterization and Global Proteomics written by Jeffrey Clark Joyner and published by . This book was released on 2006 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Abstract: Proteins occupy more than 50% of the dry weight of the average human cell. The proteins present in a given cell at a given time reflect the function and specialization of that cell. In addition to the specialization of cells within the body leading to various normal protein states, protein 'sub-states' can be induced by various conditions such as infections or cancer [9,21]. These disease-induced protein sub-states result in both qualitative and quantitative differences in both ribosomal production of proteins and posttranslational modification of proteins [2]. Each disease-induced sub-state therefore has its own protein profile or signature. For diseases such as cancer, which can easily progress undetected and whose manifestations have grave consequences, statistical comparisons of a patient's protein profile with many cancer-state protein profiles can be made for early detection. Two-dimensional-liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) holds much promise in the analysis of such diseaseinduced protein profiles. 2D-LC-MS/MS uses two means of separation prior to online mass analysis, whereas one-dimensional (1D) methods use only one means of separation. The increased separation of a 2D system allows for much more complete mass analysis of proteins and results in the identification and characterization of many more proteins and modifications than with a 1D system [15]. By comparing patients' protein profiles with a comprehensive proteomic database, 2D-LC-MS/MS could serve as the critical step in an efficient early-detection method for diseases such as cancer. Before protein profiles can be accurately compared and linked with diseases, a standardized technique must be used, and mechanisms that account for experimental variations must be implemented. The 2DLC- MS/MS technique proposed here is both very useful and can be standardized easily.
Download or read book LC MS MS in Proteomics written by Pedro R. Cutillas and published by Humana Press. This book was released on 2011-04-06 with total page 330 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.
Download or read book Proteome Research written by Marc R. Wilkins and published by Springer Science & Business Media. This book was released on 1997 with total page 270 pages. Available in PDF, EPUB and Kindle. Book excerpt: Introduction to the proteome (K. L. Williams, D. F. Hochstrasser). Two-dimensional electrophoresis: the state of the art and future directions (B. R. Herbert, J.-C. Sanchez, L. Bini). large-scale comparative protein modeling ( M. C. Peitsch, N. Guex); Clinical and biomedical applications of proteomics (D. F. Hochstrasser). Biological applications of proteomics (K. L. Williams, V. Pallini). Conclusions (D. F. Hochstrasser, L. Williams). Index.
Download or read book Application of Liquid Chromatography mass Spectrometry based Protein and Proteomic Analytical Approaches to Chinese Hamster Ovary Cell Based Industrial Biopharmaceutical Production written by Yuanwei Gao and published by . This book was released on 2016 with total page 210 pages. Available in PDF, EPUB and Kindle. Book excerpt: Therapeutic proteins have emerged rapidly over the past several decades, providing effective and innovative medicines for a wide range of previously refractory human diseases. Chinese hamster ovary (CHO) cells have become the predominant choice as the cellular expression system for such therapeutic production in the biopharmaceutical industry. The high throughput of the protein drug production depends on both the efficient upstream process yielding high product titers and proficient downstream purification with high product recovery and effective impurity removal. Numerous efforts have been made at both of the up- and down-stream processes of CHO-based manufacturing to improve productivity. Although advances have been achieved, many challenges remain. The underlying biology of CHO cell productivity has not been fully understood due to an incomplete biological picture, hampering the efforts of cell cultivation optimization. Moreover, it is challenging to apply the results of cell cultivation development received from the bench-top scale to large scale production bioreactors, since different behaviors of the CHO cell are frequently observed with different bioreactor types and sizes. At the same time, efficient downstream purification is also essential to ensure drug product quality. Considering the potential safety risks to patients, the identification and quantitation of impurity residues in therapeutic proteins, especially host cell proteins (HCP), is of great importance but challenging due to the bulk drug product background. New analytical technologies and strategies which can be applied to the therapeutic protein production process are needed. Liquid chromatography-mass spectrometry (LC-MS)-based approaches are a powerful tool for proteomics and protein analysis, capable of providing the most comprehensive information to date. LC-MS analysis has been extending the depth and accuracy of proteomics study. Global cell constituent analysis or 'Omics, including proteomics and metabolomics, can provide in depth global characterization of CHO cells. A deeper understanding of CHO biology can potentially improve the optimization of manufacturing bioprocesses. Moreover, LC-MS-based methods are also a great candidate for HCP analysis. This dissertation aims at adapting state-of-the art LC-MS-based protein and proteomic approaches to the industrial biopharmaceutical processes, for the benefit of industrial therapeutic drug production. In Chapter 1, the industrial therapeutic protein production platform is introduced as well as the technology of LC-MS-based protein and proteomics analysis. In Chapter 2, a study is presented where a CHO-DG44 production cell line showed different phenotypic behaviors during the scaling-up process when cultured in the production scale (5-KL scale) and bench-top scale (20-L) bioreactors with two copper levels in the culture media for each scale. Relative quantitative proteomics based on high-resolution two dimensional liquid chromatography coupled to tandem mass spectrometry (2D-LC-MS/MS) was applied. Multi-omics including proteomics and metabolomics were employed to study CHO cell systems in order to understand the phenotypic behavior. The results revealed that CHO cells underwent intermittent hypoxia in the large production bioreactor due to the less efficient oxygen transfer and longer mixing times compared to the bench-top scale. This resulted in lower productivity and viability for the production scale. In collaboration with Simion Kreimer, Ph.D. candidate in chemistry at Northeastern, Chapter 3 describes a workflow of HCP analysis in a therapeutic monoclonal antibody, taking the advantage of the high resolution capabilities of the Orbitrap mass spectrometer. A spectral library was developed based on two-dimensional high pH/low pH reversed phase (RP/RP) liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) with data dependent acquisition (DDA). Then, a novel data independent acquisition-to- parallel reaction monitoring (DIA-to-PRM) approach was developed for HCP identification and quantitative estimation. The methodology is demonstrated to be capable of detecting HCPs at the low ppm level in the bulk product background after purification. Several HCPs were quantified with isotopically labeled peptides as internal standards. The studies described in this dissertation demonstrate the power of LC-MS-based approaches to address biopharmaceutical industry needs, by studying CHO biology as well as evaluating impurities in final product. In future studies, the discovery and method developed in this thesis can be applied to improve biopharmaceutical productivity and quality.
Download or read book Proteome Analysis written by David W Speicher and published by Elsevier. This book was released on 2004-05-18 with total page 394 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book explores the current status of proteomics, an exciting new discipline, which is less than 10 years old. This new field has rapidly grown into a major commercial and research enterprise with great prospects for dramatically advancing our knowledge of basic biological and disease processes. The contributors to this book are an international panel of proteomics experts, who review and discuss the current status of specific technologies and approaches. Proteomics represents an exciting new way to pursue biological and biomedical science at an unprecedented pace. Proteomics takes a broad, comprehensive, systematic approach to understanding biology that is generally unbiased and not dependent upon existing knowledge. The major components of proteomics from basic discovery using a range of alternative analytical methods to discovery validation and use for clinical applications are discussed. State-of-the-art protein profiling methods include high resolution two-dimensional gels, two-dimensional differential in-gel electrophoresis, LC-MS and LC-MS/MS using accurate mass tags, and protein identifications of proteins from gels using mass spectrometry methods are discussed in depth. Other chapters describe comprehensive characterization of proteomes using electrophoretic prefractionation and analyses of sub-proteomes based on specific posttranslational modifications including the phospho-proteome, the glyco-proteome, and nitrated proteins. These conventional proteome analysis chapters are complemented by discussion of emerging technologies and approaches such as affinity based biosensor proteomics as well as the use of protein microarrays, microfluidics and nanotechnology. Strategies for improving throughput by automation are also discussed. Additional chapters address the application of current proteome techniques to clinical problems and the availability of protein expression library resources for proteome studies. · Authored by international experts in the field · Covers a wide range of topics including 2-D gels, global proteomics using accurate mass tags, global proteomics using electrophoretic prefractionation, microfluidics, and nanotechnology· Includes state-of-the-art protein profiling methods, and emerging technologies
Download or read book Multidimensional Liquid Chromatography written by Steven A. Cohen and published by Wiley-Interscience. This book was released on 2008-04-04 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Multidimensional Liquid Chromatography (MDLC) is a very powerful separation technique for analyzing exceptionally complex samples in one step. This authoritative reference presents a number of recent contributions that help define the current art and science of MDLC. Topics covered include instrumentation, theory, methods development, and applications of MDLC in the life sciences and in industrial chemistry. With the information to help you perform very difficult separations of complex samples, this reference includes chapters contributed by leading experts or teams of experts.
Download or read book The Expanding Role of Mass Spectrometry in Biotechnology written by Gary Siuzdak and published by . This book was released on 2006 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covers the basic concepts in mass spectrometry as well as advanced topics including protein identification/protein structural analysis, carbohydrate and oligonucleotide analysis. Topics also include pharmacokinetics, high throughput screening, and the recent development of mass spectrometry in clinical diagnosis.
Download or read book Principles and Applications of Clinical Mass Spectrometry written by Nader Rifai and published by Elsevier. This book was released on 2018-06-26 with total page 218 pages. Available in PDF, EPUB and Kindle. Book excerpt: Principles and Applications of Clinical Mass Spectrometry: Small Molecules, Peptides, and Pathogens is a concise resource for quick implementation of mass spectrometry methods in clinical laboratory work. Focusing on the practical use of these techniques, the first half of the book covers principles of chromatographic separations, principles and types of mass spectrometers, and sample preparation for analysis; the second half outlines the main applications of this technology within clinical laboratory settings, including determination of small molecules and peptides, as well as pathogen identification. A thorough yet succinct guide to using mass spectrometry technology in the clinical laboratory, Principles and Applications of Clinical Mass Spectrometry: Small Molecules, Peptides, and Pathogens is an essential resource for chemists, pharmaceutical and biotech researchers, certain government agencies, and standardization groups. Provides concrete examples of the main applications of mass spectrometry technology Describes current capabilities of the LC- and MS-based analytical methods Details methods for successful analytical work in the field
Download or read book Environmental Health Perspectives written by and published by . This book was released on 2004 with total page 1252 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Advances in the Use of Liquid Chromatography Mass Spectrometry LC MS Instrumentation Developments and Applications written by and published by Elsevier. This book was released on 2018-01-02 with total page 448 pages. Available in PDF, EPUB and Kindle. Book excerpt: Advances in the Use of Liquid Chromatography Mass Spectrometry (LC-MS): Instrumentation Developments and Application, Volume 79, highlights the most recent LC-MS evolutions through a series of contributions by world renowned scientists that will lead the readers through the most recent innovations in the field and their possible applications. Many authoritative books on LC-MS are already present in market, describing in detail the different interfaces and their principles of operation. This book focuses more on new trends, starting with the innovations of each technique, to the most progressive challenges of LC-MS. Presents an understanding of the new advancements in LC and MS which are essential for a step forward in LC-MS applications Provides insight into the state-of-the-art in the currently available LC-MS interfaces and their principle of use Expounds on the new frontiers in LC-MS and their application potential
Download or read book Biological Mass Spectrometry written by A.L. Burlingame and published by Gulf Professional Publishing. This book was released on 2005-11-28 with total page 520 pages. Available in PDF, EPUB and Kindle. Book excerpt: Describes and integrates the techniques of many advances in both chromatographic and mass spectrometric technologies. This book also covers various biophysical applications, such as H/D exchange for study of conformations, protein-protein and protein-metal and ligand interactions. It also describes atto-to-zepto-mole quantitation of 14C and 3H.
Download or read book Quantitative Proteome Analysis written by Kazuhiro Imai and published by CRC Press. This book was released on 2013-08-05 with total page 314 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book focuses on the advantages and disadvantages of each of the commonly used quantitative proteomic methods in terms of accuracy, sensitivity, and reproducibility. It also concentrates on the effective applications of these methods that resulted in many discoveries of the role of the proteins expressed in living cells and biological fluids. The first part of the book focuses on the description of advantages and disadvantages of each of the commonly used quantitative proteomic methods in terms of accuracy, sensitivity, and, especially, reproducibility. The second part of the book focuses on providing concise descriptions of the effective applications of these methods to demonstrate how they have resulted in many important discoveries of the roles of the proteins expressed in living cells.
Download or read book Proteome Analysis written by David W Speicher and published by Elsevier. This book was released on 2004-03-26 with total page 398 pages. Available in PDF, EPUB and Kindle. Book excerpt: State-of-the-art protein profiling methods include high resolution two-dimensional gels, two-dimensional differential in-gel electrophoresis, LC-MS and LC-MS/MS using accurate mass tags, and protein identifications of proteins from gels using mass spectrometry methods are discussed in depth. Other chapters describe comprehensive characterization of proteomes using electrophoretic prefractionation and analyses of sub-proteomes based on specific posttranslational modifications including the phospho-proteome, the glyco-proteome, and nitrated proteins. These conventional proteome analysis chapters are complemented by discussion of emerging technologies and approaches such as affinity based biosensor proteomics as well as the use of protein microarrays, microfluidics and nanotechnology. Strategies for improving throughput by automation are also discussed.-
Download or read book Proteomics Sample Preparation written by Jörg von Hagen and published by John Wiley & Sons. This book was released on 2011-08-24 with total page 498 pages. Available in PDF, EPUB and Kindle. Book excerpt: This long-awaited first guide to sample preparation for proteomics studies overcomes a major bottleneck in this fast growing technique within the molecular life sciences. By addressing the topic from three different angles -- sample, method and aim of the study -- this practical reference has something for every proteomics researcher. Following an introduction to the field, the book looks at sample preparation for specific techniques and applications and finishes with a section on the preparation of sample types. For each method described, a summary of the pros and cons is given, as well as step-by-step protocols adaptable to any specific proteome analysis task.
Download or read book Handbook of Microdialysis written by and published by Elsevier. This book was released on 2007-02-02 with total page 713 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microdialysis is currently one of the most important in vivo sampling methods in physiology and pharmacology. It is used to determine the chemical components of the fluid in the extracellular space of tissues. The technique is now well established in neuroscience research and is used excessively in behavioral neuroscience to determine the concentrations and identities of molecules in brain tissues, and their change due to behavior, hormonal and transmitter changes in the nervous system. The book provides a detailed comprehensive overview of the technology and its applications, including application in pathology, drug development, and the application in the clinic. The authors are all well known researchers in Neuroscience and experts in the use of Microdialysis. Organized into two parts of seven sections, the Handbook of Microdialysis critically examines recent developments in the field through a variety of chapters written by an internationally acclaimed group of authors. It is the first comprehensive handbook covering the technology of Microdialysis and its applications in Neuroscience. * Presents microdialysis methods and interpretation including the technical aspects of microdialysis as a sampling technique followed by the analytical chemical methods* Discusses the role of microdialysis in pharmacology, drug development and models of CNS pathology* Includes clinical applications of microdialysis
Download or read book Immune Biology of Allogeneic Hematopoietic Stem Cell Transplantation written by Gerard Socie and published by Academic Press. This book was released on 2018-11-22 with total page 430 pages. Available in PDF, EPUB and Kindle. Book excerpt: Immune Biology of Allogeneic Hematopoietic Stem Cell Transplantation: Models in Discovery and Translation, Second Edition once again provides clinical and scientific researchers with a deep understanding of the current research in this field and the implications for translational practice. By providing an overview of the immune biology of HSCT, an explanation of immune rejection, and detail on antigens and their role in HSCT success, this book embraces biologists and clinicians who need a broad view of the deeply complex processes involved. It then moves on to discuss the immunobiology mechanisms that influence graft-versus-host disease (GVHD), graft-versus-leukemia effect, and transplantation success. Using illustrative figures, highlighting key issues, describing recent successes, and discussing unanswered questions, this book sums up the current state of HSCT to enhance the prospects for the future. The second edition is fully revised and includes new chapters on microbiome, metabolism, kinase targets, micro-RNA and mRNA regulatory mechanisms, signaling pathways in GVHD, innate lymphoid system development, recovery and function in GVHD, genetically engineered T-cell therapies, immune system engagers for GVHD and graft-versus-tumor, and hematopoietic cell transplant for tolerance induction in solid organ grafts. Brings together perspectives from leading laboratories and clinical research groups to highlight advances from bench to the bedside Guides readers through the caveats that must be considered when drawing conclusions from studies with animal models before correlating to clinical allogeneic hematopoietic stem cell transplantation (HSCT) scenarios Categorizes the published advances in various aspects of immune biology of allogeneic HSCT to illustrate opportunities for clinical applications
Download or read book Chemical Proteomics written by Gerard Drewes and published by Humana Press. This book was released on 2011-11-08 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The multidisciplinary science of chemical proteomics studies how small molecules of synthetic or natural origin bind to proteins and modulate their function. In Chemical Proteomics: Methods and Protocols, expert researchers in the field provide key techniques to investigate chemical proteomics focusing on analytical strategies, how probes are generated, techniques for the discovery of small molecule targets and the probing of target function, and small molecule ligand and drug discovery. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Chemical Proteomics : Methods and Protocols seeks to provide methodologies that will contribute to a wider application of chemical proteomics methods in biochemical and cell biological laboratories.
