Antibodies are indispensable tools for research, diagnosis, and therapy. Recombinant approaches allow the modification and improvement of nearly all antibody properties, such as affinity, valency, specificity, stability, serum half-life, effector functions, and immunogenicity.
Antibody Engineering provides a comprehensive toolbox covering the well-established basics but also many exciting new techniques. The protocols reflect the latest "hands on" knowledge of key laboratories in this still fast-moving field. Newcomers will benefit from the proven step-by-step protocols, which include helpful practical advice; experienced antibody engineers will appreciate the new ideas and approaches. The book is an invaluable resource for all those engaged in antibody research and development.
Inhalt
Bioinformatics of Antigen-binding Sites.- Analysis of Single Chain Antibody Sequences Using the VBASE2 Fab Analysis Tool.- Standardized Sequence and Structure Analysis of Antibody Using IMGT®.- Protein Sequence and Structure Analysis of Antibody Variable Domains.- Generation of Antibody Fragments and Their Derivatives.- scFv by Two-Step Cloning.- Bivalent Diabodies.- Generation of Single-Chain Fv Fragments and Multivalent Derivatives scFv-Fc and scFv-CH3 (Minibodies).- Miniantibodies.- Generation of Stably Transfected Eukaryotic Cell Lines Producing ImmunoRNAse Fusion Proteins.- AntibodyCytokine Fusion Proteins with Members of the TNF-Family.- Recombinant Immunotoxins for Treating Cancer.- T Bodies.- Expressing Intrabodies in Mammalian Cells.- Phenotypic Knockdown with Intrabodies.- Disulfide-Stabilized Fv Fragments.- PEGylation of Antibody Fragments to Improve Pharmacodynamics and Pharmacokinetics.- Fusion Proteins with Improved PK.- In Vivo Biotinylated scFv Fragments.- Bispecific Diabodies and Single-Chain Diabodies.- Generation and Characterization of a Dual Variable Domain Immunoglobulin (DVD-Ig™) Molecule.- Isolation of Antigen-Specific Nanobodies.- CDR-FR Peptides.- Production of Antibody Fragments.- Purification and Characterization of His-Tagged Antibody Fragments.- Production of Antibody Fragments in the Gram-Positive Bacterium Bacillus megaterium.- Analysis and Purification of Antibody Fragments Using Protein A, Protein G, and Protein L.- Purification and Analysis of Strep-tagged Antibody-Fragments.- Production of Antibodies and Antibody Fragments in Escherichia coli.- Improving Expression of scFv Fragments by Co-expression of Periplasmic Chaperones.- Bioreactor Production of scFv Fragments in Pichia pastoris.- Expression of Antibody Fragments in Transgenic Plants.- Transient Production of scFv-Fc Fusion Proteins in Mammalian Cells.- Recombinant Antibody Molecules in Nanobiotechnology and Proteomics.- Immunoliposomes.- Targeted Polymeric Nanoparticles.-Antibody Microarrays for Expression Analysis.- Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple Spotting Technique.- Preclinical and Clinical Development.- Xenograft Mouse Models for Tumour Targeting.- Xenograft Mouse Models for Tumour Targeting.- Imaging Tumor Xenografts Using Radiolabeled Antibodies.- Human Anti-antibody Response.- IP Issues in the Therapeutic Antibody Industry.