Protein Chromatography

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.

With its focus on process development and large-scale bioseparation tasks, this is tailor-made reading for the professional bioengineer in both the biotech and pharmaceutical industries. Following a tried-and-tested concept, this guide has been developed over several years in training courses for biotech and chemical engineers in Europe and the U.S. The first part deals with the theory, introducing chromatography and its dynamics, as well as discussing mass transfer and dispersion effects. The second part then goes on to cover equipment and protocols, determining the retention factor and HETP from isocratic and elution experiments, as well as the mass transfer and intraparticle diffusivity from batch and shallow-bed adsorption experiments.

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.

New textbooks at alllevels of chemistry appear with great regularity. Some fields like basic biochemistry, organic reaction mechanisms, and chemical thermodynamics are weil represented by many excellent texts, and new or revised editions are published sufficiently often to keep up with progress in research. However, some areas of chemistry, especially many of those taught at the graduate level, suffer from a reallack ofup-to-date textbooks. The most serious needs occur in fields that are rapidly changing. Textbooks in these subjects usually have to be written by scientists actually involved in the research which is advancing the field. lt is not often easy to persuade such individuals to settime aside to help spread the knowledge they have accumu lated. Our goal, in this series, is to pinpoint areas of chemistry where recent progress has outpaced what is covered in any available textbooks, and then seek out and persuade experts in these fields to produce relatively concise but instructive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In some cases the availability of texts in active research areas should help stimulate the creation of new courses. NewYork CHARLES R. CANTOR Preface to the Second Edition The original plan for the first edition of this book was to title it Enzyme Purification: Princip/es and Practice.

The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.

New textbooks at all levels of chemistry appear with great regularity. Some fields such as basic biochemistry, organic re action mechanisms, and chemical thermodynamics are weil represented by many excellent texts, and new or revised editions are published sufficiently often to keep up with progress in research. However, some areas of chemistry, especially many of those taught at the graduate level, suffer from a real lack of up-to-date textbooks. The most serious needs occur in fields that are rapidly changing. Textbooks in these subjects usually have to be written by scientists actually involved in the research that is advancing the field. It is not often easy to persuade such individuals to set time aside to help spread the knowledge they have accumulated. Our goal, in this series, is to pinpoint areas of chemistry where recent progress has outpaced what is covered in any available textbooks, and then seek out and persuade experts in these fields to produce relatively concise but instruc tive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In so me cases the availability of texts in active research areas should help stimulate the creation of new courses.

Protein Purification provides a guide to the major techniques, including non-affinity absorption techniques, affinity procedures, non-absorption techniques and methods for monitoring protein purity. There is an overview of protein strategy and equipment, followed by discussions and examples of each technique and its applications. The basic theory and simple explanations given in Protein Purification make it an ideal handbook for final year undergraduates, and postgraduates, who are conducting research projects. It will also be a useful guide to more experienced researchers who need a good overview of the techniques and products used in protein purification.