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Throughout the more than 20 years that have followed the beginnings of capillary electrophoresis (CE), its application to the analysis of proteins and peptides has continued to be reliable, versatile, and productive. Over time, CE has matured to become a superb complement to HLPC, and in many cases has also evolved as an automated and quantitative replacement for conventional slab gel electrophoresis methods such as SDS-PAGE and isoelectric focusing. Within Capillary Electrophoresis of Proteins and Peptides, we have assembled contributions from researchers who are applying state-of-the-art CE for protein and peptide analysis, including topics that we believe are of great potential both in the present and for the future. In comparison to traditional separation methods, CE represents a miniaturized analysis technique (especially in its microchip-based format) that is highly dependent upon the basic fundamentals of effective sample recovery and high sensitivity detection. With these issues in mind, Chapters 1–4 describe recently developed approaches for both capillary coatings and analyte detection via laser-induced fluorescence. Since the discipline of biotechnology has established itself as a primary platform for the application of CE to the analysis of proteins and peptides, Chapters 5–7 demonstrate a variety of examples of the specific techniques that have been applied for the development of biopharmaceuticals and their commercialization. The methods covered here include also the analysis of oligosaccharides from glycoproteins.
This new book on capillary electrophoresis (CE) is unique in its focus on biotechnology. It is devoted to proteins, peptides, and techniques especially useful in the area of recombinant DNA products. Emphasis is also placed on glycoproteins. Because of the growing role of the glycosylation process in CE, a comprehensive chapter on the subject acts as a book within a book. Although this well-known researcher in biotechnology presents a number of chapters extensively discussing theories, important practical aspects in the routine use of capillary electrophoresis are also covered.
Over the last decade, high performance Capillary electrophoresis (HPCE) has emerged as a powerful and versatile separation technique that promises to rival high performance liquid chromatography when applied to the separation of both charged and neutral species. The high speed and high separation efficiency which can be attained using any of the various modes of HPCE has resulted in the increased use of the technique in a range of analytical environments. The procedures are, however, still in the early stages of development and several barriers remain to their adoption as the technique of choice for a range of analytical problems. One such barrier is the selection and optimization of the conditions required to achieve reproducible separations of analytes and it is in this area that this new book seeks to give assistance. The book is written by an international team of authors, drawn from both academic and industrial users, and the manufacturers of instruments. At its heart are a number of tables, divided into specific application areas. These give details of published separations of a wide range of archetypal analytes, the successful separation conditions and the matrix in which they were presented. These tables are based on separations reported since 1992 and are fully referenced to the original literature. The tables are supported by discussions of the problems that a particular area presents and the strategies and solutions adopted to overcome them. The general areas covered are biochemistry, pharmaceutical science, bioscience, ion analysis, food analysis and environmental science.
High performance liquid chromatography (HPLC) is a technique used in diverse laboratory and industrial settings for the separation of components of complex mixtures. HPLC: Practical and Industrial Applications is a trouble-shooting, problem-solving guide for scientists, engineers, and technicians who use HPLC in their day-to-day work. It provides the answers to specific problems and includes practical case studies. This case history approach to chromatography is an effective teaching tool and clearly illustrates how to use techniques such as reversed phase chromatography, ion exchange chromatography, gel permeation chromatography, and capillary electrophoresis. The book is organized to facilitate rapid understanding for those working with a particular area of chromatography: Introduces the reader to instrumentation, reviews basic chromatographic theory, and presents a brief survey of absorbance fluorescence and refractive index detectors. Provides a broad view of the role of the analytical laboratory in an industrial organization. Offers suggestions on optimizing the utilization of personnel and work flow in the laboratory. Covers process sampling and analysis · Describes process chromatography. The remaining chapters discuss specialties within separations technology, including an outline of the key features of each technique, a thorough bibliography, a list of precedents, and detailed examples of one or more applications presented from the viewpoint of industrial and basic scientists. Specialty detectors are also described. HPLC: Practical and Industrial Applications is an essential reference for those working in the industrial sector, as well as scientists, students, technicians interested in learning HPLC methods.
Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. - Covers all theoretical and practical aspects of capillary gel electrophoresis - Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods - Teaches readers how to use the technology and select methods that are ideal for fundamental problems - Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
In the 1980s, capillary electrophoresis (CE) joined high-performance liquid chromatography (HPLC) as the most powerful separation technique available to analytical chemists and biochemists. Published research using CE grew from 48 papers in the year of commercial introduction (1988) to 1200 in 1997. While only a dozen major pharmaceutical and biotech companies have reduced CE to routine practice, the applications market is showing real or potential growth in key areas, particularly in the DNA marketplace for genomic mapping and forensic identification. For drug development involving small molecules (including chiral separations), one CE instrument can replace 10 liquid chromatographs in terms of speed of analysis. CE also uses aqueous rather than organic solvents and is thus environmentally friendlier than HPLC. The second edition of Practical Capillary Electrophoresis has been extensively reorganized and rewritten to reflect modern usage in the field, with an emphasis on commercially available apparatus and reagents. This authoritative and very comprehensible treatment builds on the author's extensive experience as an instructor of short courses for the American Chemical Society and for industry. - Illustrated with detailed diagrams of electrophoretic phenomena - Offers step-by-step methods development schemes - Presents techniques for developing quantitative, robust, and precise methods - Includes an extensive troubleshooting guide - Updates and greatly expands on the first edition-more than 50% of the text is new - Written by an internationally recognized scientist who is an instructor for American Chemical Society short courses on HPCE
Furthering efforts to simulate the potency and specificity exhibited by peptides and proteins in healthy cells, this remarkable reference supplies pharmaceutical scientists with a wealth of techniques for tapping the enormous therapeutic potential of these molecules-providing a solid basis of knowledge for new drug design. Provides a broad, comprehensive overview of peptides and proteins as mediators of cell movement, proliferation, differentiation, and communication. Written by more than 50 leading international authorities, Peptides and Protein Drug Analysis discusses strategies for dealing with the complexity of peptides and proteins in conformational flexibility and amino acid sequence variability analyzes drug formulations facilitated by solid-phase peptide synthesis and recombinant DNA technology examines chemical purity analysis by high-pressure chromatographic, capillary electrophoretic, gel electrophoretic, and isoelectric focusing methods highlights drug design elements derived from protein folding, bioinformatics, and computational chemistry demonstrates uses of unnatural mutagenesis and combinatorial chemistry explores mass spectrometry, protein sequence, and carbohydrate analysis illustrates bioassays and other new functional analysis methods surveys spectroscopic techniques such as ultraviolet, fluorescence, Fourier transform infrared, and nuclear magnetic resonance (NMR) addresses ways of distinguishing between levels of therapeutic and endogenous agents in cells reviews structural analysis tools such as ultracentrifugation and light, X-ray, and neutron scattering and more! Featuring over 3400 bibliographic citations and more than 500 tables, equations, and illustrations, Peptide and Protein Drug Analysis is a must-read resource for pharmacists; pharmacologists; analytical, organic, and pharmaceutical chemists; cell and molecular biologists; biochemists; and upper-level undergraduate and graduate students in these disciplines.
Capillary Electrophoresis (CE) has had a very significant impact on the field of analytical chemistry in recent years as the technique is capable of very high resolution separations, requiring only small amounts of samples and reagents. Furthermore, it can be readily adapted to automatic sample handling and real time data processing. Many new methodologies based on CE have been reported. Rapid, reproducible separations of extremely small amounts of chemicals and biochemicals, including peptides, proteins, nucleotides, DNA, enantiomers, carbohydrates, vitamins, inorganic ions, pharmaceuticals and environmental pollutants have been demonstrated. A wide range of applications have been developed in greatly diverse fields, such as chemical, biotechnological, environmental and pharmaceutical analysis.This book covers all aspects of CE, from the principles and technical aspects to the most important applications. It is intended to meet the growing need for a thorough and balanced treatment of CE. The book will serve as a comprehensive reference work. Both the experienced analyst and the newcomer will find the text useful.
Because new information was discovered at an incredible rate since the publication of the successful first edition of this Handbook, this fully updated second edition covers all areas of interest in the field of capillary electrophoresis (CE). A relatively new technology, CE is a principle method for studying the physicochemical properties of proteins, peptides, and other macromolecules. Where applicable, the 30 chapters provide basic underlying theories as well as application-oriented aspects of each technique.Keep up with all the developments in this growing field with the Handbook of Capillary Electrophoresis, Second Edition - a complete guide to the fundamentals of CE and the latest research. The chapters are organized into five units: Modes: Presents a theoretical development of the basic principles governing separation with several modes, including CEC, and discusses their practical aspects. Analyte: Applies CE to the analysis of a specific class of analytes, including organic and inorganic ions, pharmaceuticals, glycoconjugates, peptides, proteins, and DNA fragments. Fundamental Aspects of CE: Technique-oriented information for the practitioner, including the importance of the sample matrix, on-line preconcentration of samples, modes of detection, and specific aspects of CE data analysis. Applications of CE: Includes single cell analysis, CE in DNA sequencing, CE as a clinical diagnostic tool, identifying and quantifying drugs, and for characterizing interacting species. Specialized Aspects of CE: Discusses interfacing CE with mass spectrometry, high-volume throughput continuous CE, microchip CE, control of EOF, and much more. The Handbook of Capillary Electrophoresis, Second Edition, pulls together diverse areas and applications of CE, resulting in an excellent tool for scientists involved in biotechnology and clinical chemistry, as well as the pharmaceutical, bioscience, chemical, and instrument-manufacturing industries. With an applications-oriented focus, the handbook is also a superb manual for workshops, seminars, and graduate courses in separation science.