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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. Authoritative and up-to-date, Antibody Engineering: Methods and Protocols, Third Edition presents the reader with an extensive toolbox to create the powerful molecules of tomorrow.
In Silico Methods
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
Generation of Diversity
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
Selections of Lead Candidate
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
Production of Recombinant Antibodies
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
Fc Engineering
Chemoenzymatic Defucosylation of Therapeutic Antibodies for Enhanced Effector Functions Using Bacterial α-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
Characterization, Optimization, and Innovative Formats
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