This book is a collection of critical reviews of the use of high-performance liquid chromatography in a very specialized area of research. It describes in detail modern methodology to separate nucleic acids, enzymes and a wide variety of biologically active proteins such as renin.
Specialist Periodical Reports provide systematic and detailed review coverage of progress in the major areas of chemical research. Written by experts in their specialist fields the series creates a unique service for the active research chemist, supplying regular critical in-depth accounts of progress in particular areas of chemistry. For over 80 years the Royal Society of Chemistry and its predecessor, the Chemical Society, have been publishing reports charting developments in chemistry, which originally took the form of Annual Reports. However, by 1967 the whole spectrum of chemistry could no longer be contained within one volume and the series Specialist Periodical Reports was born. The Annual Reports themselves still existed but were divided into two, and subsequently three, volumes covering Inorganic, Organic and Physical Chemistry. For more general coverage of the highlights in chemistry they remain a 'must'. Since that time the SPR series has altered according to the fluctuating degree of activity in various fields of chemistry. Some titles have remained unchanged, while others have altered their emphasis along with their titles; some have been combined under a new name whereas others have had to be discontinued. The current list of Specialist Periodical Reports can be seen on the inside flap of this volume.
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.
Current Research in Protein Chemistry: Techniques, Structure, and Function focuses on the techniques and methods used for determining the structure and function of proteins. Topics covered range from protein folding and stability to catalysis by chimeric proteins, amino acid and peptide analysis, applications of mass spectrometry to peptide and protein analysis, and protein sequencing. This book is divided into six sections encompassing 55 chapters. The first chapter describes a novel method for protein hydrolysis by means of microwave irradiation that uses Teflon-Pyrex tubes. This is followed by a discussion of the application of high performance capillary electrophoresis to the analysis of amino acids. The sections that follow focus on mass spectrometric methods, protein sequencing, and capillary electrophoresis as well as protein stability, chimeric proteins and enzyme modifications, and protein structure prediction. The crystal structure of human interleukin-1alpha, the acid-denatured states of proteins, solubility of recombinant proteins expressed in Escherichia coli, and catalysis by chimeric proteins are considered. The reader is also introduced to peptide mapping and internal sequencing of proteins from acrylamide gels, new approaches to covalent sequence analysis, alkaline denaturation of hemoglobin, and measurements of disulfide bond stabilities in protein folding intermediates. Students and researchers interested in protein chemistry will find this book extremely helpful.
This book covers the fundamentals of protein inactivation during bioseparation and the effect on protein processing. Bioseparation of Proteins is unique because it provides a background of the bioseparation processes, and it is the first book available to emphasize the influence of the different bioseparation processes on protein inactivation.Bioseparation of Proteins covers the extent, mechanisms of, and control of protein inactivation during these processes along with the subsequent and essential validation of these processes. The book focuses on the avoidance of protein (biologicalproduct) inactivation at each step in a bioprocess. It compares protein inactivation exhibited during the different bioseparation processes by different workers and provides a valuable framework for workers in different areas interested in bioseparations.Topics include separation and detection methods; estimates of protein inactivation and an analysis of this problem for different separation processes; strategies for avoiding inactivation; the molecular basis of surface activity and protein adsorption,process monitoring, and product validation techniques; and the economics of various bioseparation processes and quality control procedures.Key Features* Protein inactivation and other aspects of biological stability are critical to an effective bioseparation process; This book is a detailed and critical review of the available literature in an area that is essential to the effectiveness, validation, and economics of bioseparation processes for drugs and other biological products; Conveniently assembled under one cover, the survey of the literature and resulting perspective will greatly assist engineers and chemists in designingand improving their own processes; Key features of the text include:* detailed data on biological stability under various bioseparation conditions* extensive case studies from the literature on separation processes, validation, and economics* simplified analysis of protein refolding and inactivation mechanisms* consideration of adsorption theories and the effect of heterogeneity* coverage of both classical and novel bioseparation techniques, including chromatographic procedures
Completely revised to reflect the innovations in HPLC from the past decade, this authoritative reference presents practical strategies for the evaluation and analysis of proteins, peptides, and polynucleotides. Offering class-specific applications for the characterization and fractionation of biological macromolecules, the book contains material on organic supports, size exclusion, ion exchange, hydrophobic interaction, and metal interaction chromatography. Leading experts summarize specialized detection systems, provides discussions on the chemical and biological properties of specific biomolecules, include detailed guidelines for the development of analytical techniques, and more.