Protein Liquid Chromatography is a handbook-style guide to liquid chromatography as a tool for isolating and purifying proteins, consisting of 25 individual chapters divided into three parts: Part A covers commonly-used, classic modes of chromatography such as ion-exchange, size-exclusion, and reversed-phase; Part B deals with various target protein classes such as membrane proteins, recombinant proteins, and glycoproteins; and Part C looks at various miscellaneous related topics, including coupling reaction, buffer solution additives, and software. The text as a whole can be viewed as a systematic survey of available methods and how best to use them, but also attempts to provide an exhaustive coverage of each facet. How to solve a specific problem using a chosen method is the overall essence of the volume. The principle philosophy of this compilation is that practical application is everything; therefore, both classical and modern methods are presented in detail, with examples involving conventional, medium- and high-pressure techniques. Over-exposure to history, concept, and theory has deliberately been avoided. The reader will find a wealth of tips and tricks from users for users, including advice on the advantages and disadvantages of each method. Easy-to-read sections on "Getting started now" and "Where to go from here" attempt to provide hands-on, fool-proof detailed practical procedures with complete and even standard model runs for any scientist or technician at work in this area.

A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Protocols, leading researchers present clear protocol-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular BiologyTM series format, protocols chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Protocols will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.

An all-in-one practical guide on how to efficiently use chromatographic separation methods Based on a training course that teaches the theoretical as well as practical aspects of protein bioseparation to bioprocess professionals, this fully updated and revised new edition offers comprehensive coverage of continuous chromatography and provides readers with many relevant examples from the biopharmaceutical industry. Divided into two large parts, Protein Chromatography: Process Development and Scale-Up, Second Edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. The first part introduces chromatographic theory, including process design principles, to enable the reader to rationalize the set-up of a bioseparation process. The second part illustrates by way of case studies and sample protocols how the theory learned in the first part may be applied to real-life problems. Chapters look at: Downstream Processing of Biotechnology Products; Chromatography Media; Laboratory and Process Columns and Equipment; Adsorption Equilibrium; Rate Processes; and Dynamics of Chromatography Columns. The book closes with chapters on: Effects of Dispersion and Rate Processes on Column Performance; Gradient Elution Chromatography; and Chromatographic Column Design and Optimization. -Presents the most pertinent examples from the biopharmaceutical industry, including monoclonal antibodies -Provides an overview of the field along with design tools and examples illustrating the advantages of continuous processing in biopharmaceutical productions -Focuses on process development and large-scale bioseparation tasks, making it an ideal guide for the professional bioengineer in the biotech and pharma industries -Offers field-tested information based on decades of training courses for biotech and chemical engineers in Europe and the U.S. Protein Chromatography: Process Development and Scale-Up, Second Edition will appeal to biotechnologists, analytical chemists, chromatographers, chemical engineers, pharmaceutical industry, biotechnological industry, and biochemists.

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.

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

Hans Neurath has written that this is the second golden era of enzymology {Protein Science [1994], vol. 3, pp. 1734—1739); he could with justice have been more general and referred to the second golden age of protein chemistry. The last two decades have seen enormous advances in our understanding of the structures and functions of pro teins arising on the one hand from improvements and developments in analytical techniques {see the companion volume, Basic Protein and Peptide Protocols, in this series) and on the other hand from the tech nologies of molecular genetics. Far from turning the focus away from protein science, the ability to isolate, analyze, and express genes has increased interest in proteins as gene products. Hence, many laborato ries are now getting involved in protein isolation for the first time, either as an essential adjunct to their work in molecular genetics or because of a curiosity to know more about the products of the genes that they have been studying. Protein Purification Protocols is aimed mainly at these newcom ers to protein purification, but it is hoped that it will also be of value to established practitioners who may find here techniques that they have not tried, but which might well be most applicable in their work. With the exception mainly of the first and last chapters, the format of the contributions to the present book conform to the established format of the Methods in Molecular Biology series.

Principles and Practice of Modern Chromatographic Methods, Second Edition takes a comprehensive, unified approach in its presentation of chromatographic techniques. Like the first edition, the book provides a scientifically rigid, but easy-to-follow presentation of chromatography concepts that begins with the purpose and intent of chromatographic theory - the "what and why that are left out of other books attempting to cover these principles. This fully revised second edition brings the content up-to-date, covering recent developments in several new sections and an additional chapter on composite methods. New topics include sample profiling, sample preparation, sustainable green chemistry, 2D chromatography, miniaturization/nano-LC, HILIC, and more. - Contains thorough chapters that begin with an updated schematic overview and a visual representation of the content - Avoids the obfuscation of different terminologies and classification systems that are prevalent in the area, such as the relationship between liquid chromatography and column chromatography - Provides integrated and comprehensive topic coverage based on chromatographic bibliometrics and survey reports on the relative usage of chromatographic techniques