Antibody Engineering Methods and Protocols

This detailed new edition provides complete and easy access to a variety of antibody engineering techniques.  The volume explores topics such as the generation of native, synthetic, or immune antibody libraries, the selection of lead candidates via the different powerful and innovative display technologies, Fc engineering, as well as their production, characterization, and optimization of antibodies. Written for the highly successful Methods in Molecular Biology series, 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. 

Antibody Design and Humanization via In Silico Modeling.- Antibody Affinity Maturation by Computational Design.- Use of IMGT® Databases and Tools for Antibody Engineering and Humanization.- Construction of Human Naïve Antibody Gene Libraries.- Construction of Synthetic Antibody Libraries.- Construction of Histidine-Enriched Shark IgNAR Variable Domain Antibody Libraries for the Isolation of pH-Sensitive vNAR Fragments.- Display Technologies for Generation of Ig Single Variable Domains.- A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries.- Phage Display and Selections on Purified Antigens.- Selection of Antibodies to Transiently Expressed Membrane Proteins Using Phage Display.- Selection of Antibody Fragments against Structured DNA by Phage Display.- Selection of Antibody Fragments by Yeast Display.- Rapid Selection of High Affinity Antibody scFv Fragments Using Ribosome Display.- In Vitro Selection of Single Domain Antibody (VHH) Using cDNA Display.- Sequencing and Affinity Determination of Antigen-Specific B Lymphocytes from Peripheral Blood.- Expression of IgG Monoclonals with Engineered Immune Effector Functions.- An IRES-Mediated Tricistronic Vector for Efficient Generation of Stable, High Level Monoclonal Antibody Producing CHO DG44 Cell Lines.- Production, Purification, and Characterization of Antibody-TNF Superfamily Ligand Fusion Proteins.- Chemoenzymatic Defucosylation of Therapeutic Antibodies for Enhanced Effector Functions Using Bacterial a-Fucosidases.- Fc Glyco- and Fc Protein-Engineering: Design of Antibody Variants with Improved ADCC and CDC Activity.- Fc Engineering: Tailored Synthetic Human IgG1-Fc Repertoire for High Affinity Interaction with FcRn at pH 6.0.- Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance.- Parallel Evolution of Antibody Affinity and Thermal Stability for Optimal Biotherapeutic Development.- The Use of Somatic Hypermutation for the Affinity Maturation of Therapeutic Antibodies.- Selection and Use of Intracellular Antibodies.- Site-Specific Radioactive Labeling of Nanobodies.