High Performance Liquid Chromatography Of Peptides And Proteins

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.

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.

High-Performance Liquid Chromatography of Proteins and Peptides contains the proceedings of the first International Symposium on High-Performance Liquid Chromatography of Proteins and Peptides, held in Washington, D.C., on November 16-17, 1981. The symposium focused on the use of high-performance liquid chromatography (HPLC) in the analysis, characterization, and isolation of peptides and proteins and encompassed six sessions covering size exclusion, ion exchange, and reversed phase chromatography, as well as the use of high-performance liquid chromatography (HPLC) in protein structural studies and peptide isolation. This book is comprised of 28 chapters and begins with a discussion on the status of high-performance ion-exchange chromatography of proteins, followed by an analysis of peptic fragmentation of human immunoglobulin G using HPLC. The physicochemical basis of peptide retention with chemically bonded hydrocarbonaceous silicas and the isolation of biologically active peptides from tissue extracts are also examined. Subsequent chapters explore some additional applications of HPLC, such as cord blood screening for hemoglobin disorders; purification of commercial trypsin and chymotrypsin; characterization of human alcohol dehydrogenase isoenzymes; and structural studies of neurophysins, photolabeled derivatives, and biosynthetic precursors. This monograph should be of value to students and researchers interested in the use of HPLC to study proteins and peptides.

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.

Introduction to the general principles of HPLC; HPLC instrumentation; Mobile phases; Detection methods; Separation of free amino acids; Resolution of amino acids as diastereomeric derivatives.

All areas of high performance liquid chromatography (HPLC) relevant to the separation of amino acid and polypeptide samples are described in these comprehensive volumes. Eighty-four chapters cover the application of HPLC to research in the life sciences. Both basic information and specific details are presented on such areas as commercially available HPLC columns, ligand-exchange chromatography, peptide hormone separations, peptide antibiotic separations and drug-protein binding. Practical suggestions are given to help you avoid pitfalls which can exist in the analysis of biological materials.

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.