This third edition volume expands on the previous editions with more detailed research on the characterization of antibody antigen interactions between different users with different requirements. The chapters in this book are divided into four parts: Part One looks at the entire native antigen and covers traditional structural biology techniques such as nuclear magnetic resonance and x-ray crystallography. Part Two talks about protein fragments derived from antigens, and discusses binding regions within antigen sequence using bacterial surface display and ELISA, for example. Part Three describes the use of surface plasmon resonance spectroscopy and biolayer interferometry, and Part Four highlights methods used to identify new antigens and assess antibody cross-reactivity. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Epitope Mapping Protocols, Third Edition is a valuable resource for anyone interested in furthering their research in this expanding field.

Given the versatile utility of the determinination of epitopes, beneficial to a wide variety of scientists from immunologists to structural biologists to biotechnologists, the need for a thorough, state-of-the-art collection of experimental protocols is clear. In Epitope Mapping Protocols, Second Edition, expert contributors from a broad spectrum of scientific backgrounds update and expand the successful first edition with cutting-edge techniques and applications, including approaches to both antibody or B-cell epitope mapping and T-cell epitope mapping as well as a new section on the profiling of antibody signatures in biological fluids. Written in the popular Methods in Molecular BiologyTM series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Epitope Mapping Protocols, Second Edition is a reliable and valuable reference for all those who wish to understand and further investigate the diversifying field of epitope mapping.

Little more than three years down the line and I am already writing the Preface to a second volume to follow Protein and Peptide Analysis by Mass . What has happened in between these times to make this second venture worthwhile? New types of mass spectrometric instrumentation have appeared so that new techniques have become possible and existing techniques have become much more feasible. More particularly, however, the newer ionization te- niques, introduced for the analysis of high molecular weight materials, have now been thoroughly used and studied. As a result, there has been an en- mous improvement in the associated sample handling technology so that these methods are now routinely applied to much smaller sample amounts as well as to more intractable samples. Again, this particular community of mass spectrometry users has both increased in number and diversified. And, riding this wave of acceptance, leaders in the field have set their sights on more complex problems: molecular interaction, ion structures, quantitation, and kinetics are just a few of the newer areas reported in Mass Spectrometry of Proteins and Peptides. As with the first volume, one purpose of this collection, Mass Spectr- etry of Proteins and Peptides, is to show the reader what can be done by the application of mass spectrometry, and perhaps even to encourage the reader to venture down new paths.

Monoclonal Antibodies: Methods and Protocols, Second Edition expands upon the previous edition with current, detailed modern approaches to isolate and characterize monoclonal antibodies against carefully selected epitopes. This edition includes new chapters covering the key steps to generate high quality monoclonals via different methods, from antigen generation to epitope mapping and quality control of the purified IgG. Chapters are divided into four parts corresponding to four distinct objectives. Part I covers monoclonal antibody generation, Part II deals with monoclonal antibody expression and purification, Part III presents methods for monoclonal antibody characterization and modification, and Part IV describes selected applications of monoclonal antibodies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Monoclonal Antibodies: Methods and Protocols, Second Edition provides crucial initial steps of monoclonal antibody generation and characterization with state-of-the art protocols.

This book examines a collection of state-of-the-art methods that employ monoclonal antibodies in a clinical setting. The chapters offer in-depth description for generating mouse and recombinant humanized antibodies, and a comprehensive review of how antibodies are being used in bead-based methods for measuring proteins. This field will continue to expand and provide new and innovative techniques in the laboratory and as a basis that complements targeted therapy.

Epitope Mapping covers all the major methods for the identification and definition of epitopes. This book is intended for researchers in academia and industry involved with epitope mapping.

Glyco-engineering is being developed as a method to control the composition of carbohydrates and to enhance the pharmacological properties of monoclonal antibodies (mAbs) and other proteins. In Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols, experts in the field provide readers with production and characterization protocols of glycoproteins and glyco-engineered biopharmaceuticals with a focus on mAbs. The volume is divided in four complementary parts dealing with glyco-engineering of therapeutic proteins, glycoanalytics, glycoprotein complexes characterization, and PK/PD assays for therapeutic antibodies. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols serves as an ideal guide for scientists striving to push forward the exciting field of engineered biopharmaceuticals.

Epitope Mapping Protocols offers more than thirty protocols for antibody production and mapping-all described in detail by leading experts and illustrated by proven examples from their own work. The methods range from those that use (or can use) whole, native antigens to those that use peptides and peptide libraries and those that require antigen expressed from recombinant DNA. A method for T-cell epitopes, instructions on using "natural" variants or mutants, and techniques for producing panels of antibodies for epitope mapping are also included.

Given the versatile utility of the determinination of epitopes, beneficial to a wide variety of scientists from immunologists to structural biologists to biotechnologists, the need for a thorough, state-of-the-art collection of experimental protocols is clear. In Epitope Mapping Protocols, Second Edition, expert contributors from a broad spectrum of scientific backgrounds update and expand the successful first edition with cutting-edge techniques and applications, including approaches to both antibody or B-cell epitope mapping and T-cell epitope mapping as well as a new section on the profiling of antibody signatures in biological fluids. Written in the popular Methods in Molecular BiologyTM series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Epitope Mapping Protocols, Second Edition is a reliable and valuable reference for all those who wish to understand and further investigate the diversifying field of epitope mapping.

The introduction of monoclonal antibodies revolutionized immunology. The development of human monoclonal antibodies was inspired primarily by the enormous clinical benefits promised by these reagents which can be used as anti-inflammatory reagents, anti-tumor reagents and reagents for passive immunization in a variety of pathologies. Human Monoclonal Antibodies: Methods and Protocols presents technical protocols of cellular and molecular methods for the production, purification and application of human monoclonal antibodies, as well as review articles on related topics of human monoclonal and polyclonal antibodies. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Human Monoclonal Antibodies: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies which will prove invaluable in a clinical setting.