Single Domain Antibodies Biology Engineering And Emerging Applications

Single-domain antibodies (sdAbs) represent the minimal antigen binding-competent form of the immunoglobulin domain and have unique properties and applications. SdAbs are naturally produced as the variable domains of the heavy chain-only antibodies of camelid ruminants and cartilaginous fishes, but can also be engineered synthetically from autonomous human or mouse VH or VL domains. The scope of this research topic and associated e-book covers current understanding and new developments in (i) the biology, immunology and immunogenetics of sdAbs in camelids and cartilaginous fishes, (ii) strategies for sdAb discovery, (iii) protein engineering approaches to increase the solubility, stability and antigen-binding affinity of sdAbs and (iv) specialized applications of sdAbs in areas such diagnostics, imaging and therapeutics.

Single-domain antibodies (sdAbs) represent the minimal antigen binding-competent form of the immunoglobulin domain and have unique properties and applications. SdAbs are naturally produced as the variable domains of the heavy chain-only antibodies of camelid ruminants and cartilaginous fishes, but can also be engineered synthetically from autonomous human or mouse VH or VL domains. The scope of this research topic and associated e-book covers current understanding and new developments in (i) the biology, immunology and immunogenetics of sdAbs in camelids and cartilaginous fishes, (ii) strategies for sdAb discovery, (iii) protein engineering approaches to increase the solubility, stability and antigen-binding affinity of sdAbs and (iv) specialized applications of sdAbs in areas such diagnostics, imaging and therapeutics.

This volume covers current and emerging techniques for studying single-domain antibodies (sdAbs). Chapters guide readers through the biology and immunology of sdAbs in camelids and sharks, isolation of sdAbs, protein engineering approaches to optimize the solubility, stability, valency and antigen binding affinity of sdAbs, and specialized applications of sdAbs. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Single-Domain Antibodies: Methods and Protocols aims to be a useful, practical guide to help researchers further their studies in this field.

This highly readable textbook serves as a concise and engaging primer to the emerging field of antibody engineering and its various applications. It introduces readers to the basic science and molecular structure of antibodies, and explores how to characterize and engineer them. Readers will find an overview of the latest methods in antibody identification, improvement and biochemical engineering. Furthermore, alternative antibody formats and bispecific antibodies are discussed. The book’s content is based on lectures for the specializations “Protein Engineering” and “Medical Biotechnology” within the Master’s curriculum in “Biotechnology.” The lectures have been held at the University of Natural Resources and Life Sciences, Vienna, in cooperation with the Medical University of Vienna, since 2012 and are continuously adapted to reflect the latest developments in the field. The book addresses Master’s and PhD students in biotechnology, molecular biology and immunology, and all those who are interested in antibody engineering.

Antibody Engineering comprises in vitro selection and modification of human antibodies including humanization of mouse antibodies for therapy, diagnosis, and research. This book comprises an overview about the generation of antibody diversity and essential techniques in antibody engineering: construction of immune, naive and synthetic libraries, all available in vitro display methods, humanization by chain shuffling, affinity maturation techniques, de novo synthesis of antibody genes, colony assays for library screening, construction of scFvs from hybridomas, and purification of monoclonal antibodies by exclusion chromatography. In addition, other topics that are discussed in this book are application and mechanism of single domain antibodies, structural diversity of antibodies, immune-mediated skin reactions induced by TNF-alpha recombinant antibodies, and bioinformatic approaches to select pathogen-derived peptide sequences for antibody targets.

A valuable resource for researchers and workers in the fields of both pharmaceuticals and biotechnology as well as undergraduates in biochemistry, applied biology, biomedical sciences and pharmacy, this book compares established techniques of antibody production with the new. Antibody structure and the implications of antibody engineering are fully discussed, and a case study approach illustrates how antibodies are finding increasing use in the diagnosis and treatment of disease. The volume ends with commercial expression, purification and large-scale manufacture of antibodies and their future potential, particularly as therapeutic agents.

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