HPLC of Peptides and Proteins


Book Description

The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure through the exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide and protein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age. The design of multidimensional purification schemes to achieve high levels of product purity further demonstrates the power of HPLC techniques not only in the characterization of cellular events, but also in the production of pepti- and protein-based therapeutics. HPLC continues to be at the heart of the analytical techniques with which scientists in both academia and in industry must arm themselves to be able to fully characterize the identity, purity, and potency of peptides and proteins.




Total Chemical Synthesis of Proteins


Book Description

How to synthesize native and modified proteins in the test tube With contributions from a panel of experts representing a range of disciplines, Total Chemical Synthesis of Proteins presents a carefully curated collection of synthetic approaches and strategies for the total synthesis of native and modified proteins. Comprehensive in scope, this important reference explores the three main chemoselective ligation methods for assembling unprotected peptide segments, including native chemical ligation (NCL). It includes information on synthetic strategies for the complex polypeptides that constitute glycoproteins, sulfoproteins, and membrane proteins, as well as their characterization. In addition, important areas of application for total protein synthesis are detailed, such as protein crystallography, protein engineering, and biomedical research. The authors also discuss the synthetic challenges that remain to be addressed. This unmatched resource: Contains valuable insights from the pioneers in the field of chemical protein synthesis Presents proven synthetic approaches for a range of protein families Explores key applications of precisely controlled protein synthesis, including novel diagnostics and therapeutics Written for organic chemists, biochemists, biotechnologists, and molecular biologists, Total Chemical Synthesis of Proteins provides key knowledge for everyone venturing into the burgeoning field of protein design and synthetic biology.




High Performance Liquid Chromatography in Phytochemical Analysis


Book Description

The powerful, efficient technique of high performance liquid chromatography (HPLC) is essential to the standardization of plant-based drugs, identification of plant material, and creation of new herbal medicines. Filling the void in this critical area, High Performance Liquid Chromatography in Phytochemical Analysis is the first book to give a comp




Peptide Characterization and Application Protocols


Book Description

This book is dedicated to the characterization of peptides and their applications for the study of biochemical systems. The contributing authors are all leaders in the field of peptide research. Part I, Characterization, presents the most recent advances in select analytical techniques. Part II, Application, presents a variety of specific applications for synthetic peptides. This book is an indispensable aid in the pursuit of new directions in peptide research.




High Performance Liquid Chromatography in Plant Sciences


Book Description

Modern Methods of Plant Analysis When the handbook Modern Methods of Plant Analysis was first introduced in 1954 the considerations were: 1. the dependence of scientific progress in biology on the improvement of existing and the introduction of new methods; 2. the difficulty in finding many new analytical methods in specialized journals which are normally not accessible to experimental plant biologists; 3. the fact that in the methods sections of papers the description of methods is frequently so compact, or even sometimes so incomplete that it is difficult to reproduce experiments. These considerations still stand today. The series was highly successful, seven volumes appearing between 1956 and 1964. Since there is still today a demand for the old series, the publisher has decided to resume publication of Modern Methods of Plant Analysis. It is hoped that the New Series will be just as acceptable to those working in plant sciences and related fields as the early volumes undoubtedly were. It is difficult to single out the major reasons for success of any publication, but we believe that the methods published in the first series were up-to-date at the time and presented in a way that made description, as applied to plant material, complete in itselfwith little need to consult other publications. Contributing authors have attempted to follow these guidelines in this New Series of volumes.




Neuropeptide Protocols


Book Description

Leading scientists offer an unprecedented suite of master protocols for analyzing neuropeptide structure and function using optimized classic methods and the latest state-of-the-art techniques. This comprehensive collection includes valuable techniques for extracting and purifying neuropeptides from biological sources, for their sequence determination by Edman degradation and PCR, and for the solid-phase synthesis and purification of peptides and peptide analogs. Also covered are physical techniques for the characterization of synthetic or natural peptides, methods for immunocytochemical sample preparation and visualization at both light and electron levels, and techniques for neuropeptide-receptor interaction. Chapters on molecular modeling, radioisotopic labeling, production of antisera, the measurement of efflux from brain to blood, and the assay and inhibition of neuropeptidases complete this indispensable collection of readily reproducible protocols.




High-Performance Liquid Chromatography of Peptides and Proteins


Book Description

This book consists of a series of 82 precise, easy-to-read articles by internationally renowned scientists and emphasizes the practical approach to HPLC with minimal theory, although the underlying principles for peptide and protein separations are clearly expressed. All of the major modes of microbore, ultrafast and analytical HPLC are discussed, including size-exclusion, ion-exchange, reversed-phase, hydrophobic interaction, and affinity and immunoaffinity chromatography. A section on preparative HPLC, including displacement techniques, is also presented. Problem-solving approaches to the separation of various classes of biologically active peptides and proteins are thoroughly explored, while the importance of peptide standards for monitoring column performance and for optimizing separation conditions is emphasized. Several articles focus on the choice of the correct detection method (electrochemical, UV, fluorescence), as well as the need for a proper knowledge of approaches to column and instrument maintenance and trouble-shooting. A section on predictive approaches deals with both computer simulation of peptide separations and peptide structure. The book also includes complementary techniques to HPLC, as well as other useful applications of HPLC. It enables both novice and experienced chromatographers to realize the full potential of this extremely powerful technique, in the process making an important contribution to scientific literature.




Chapters In The Evolution Of Chromatography


Book Description

Chromatography, invented more than 100 years ago, is the most widely used separation technique in the world today. It has helped the birth of modern analytical instrumentation and continues to strongly influence the profiles of our chemical, biochemical and clinical laboratories.This book deals with the history of the invention and evolution of chromatography and of the various chromatographic techniques. After discussing the precursors, it elaborates on the activities of M.S. Tswett, the inventor of the technique, and of a few selected key pioneers. It then summarizes the evolution of the various branches of chromatography (planar, ion-exchange, gas and liquid), and also reviews the key role of international symposia in setting the trends in this evolution. Except for individual publications of the author, the history of the evolution of chromatography has not been the subject of any book. Thus, this book fills a major gap in the scientific literature./a




Protein Structure Analysis


Book Description

"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.




HPLC in Enzymatic Analysis


Book Description

The use of High Performance Liquid Chromatography (HPLC) techniques in the study of enzymatic reactions has grown significantly since the publication of the first edition of this highly successful book: the role of enzymes in biological research has expanded; the application of HPLC and enzymes has extended to more disciplines; advances in separation techniques and instrumentation have increased the capability of HPLC; and the discovery of new enzymes has spawned new methods of analysis. High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition addresses these developments in its coverage of the refinements of HPLC methods and their use in a wide range of laboratory applications. It offers the same practical approach found in the first edition, incorporates a wealth of new information into existing chapters, and adds new chapters to deal with new applications, including capillary electrophoresis, forensic chemistry, microdialysis, and the polymerase chain reaction. Topics include: * Application of HPLC to the assay of enzymatic activities * Concepts and principles of HPLC, including the latest technological advances * Concepts and principles of capillary electrophoresis (CE) * Strategy for design of an HPLC/CE system for assay of enzyme activity * Preparation of enzymatic activities from tissues and single cells * Analysis of enzymatic activities in body fluids, including chromatobiosis * HPLC for the identification of new enzymatic activities * Fundamentals of the polymerase chain reaction * HPLC in forensics * Survey of enzymatic activities assayed by the HPLC method, including many new categories * Multienzyme systems, including many new examples * HPLC in the analysis of contaminated food "It is the ability of HPLC to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently, antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of HPLC for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently." --from the preface to the First Edition Easy to read and full of practical advice and hundreds of diagrams and examples, High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition is an invaluable resource for students, researchers, and laboratory workers in analytical chemistry and biochemistry, molecular biology and cell biology, and for anyone interested in keeping up with this fast-growing field.