Description
This second edition details new and updated methods on different antibody libraries, along with novel approaches for antibody discovery. Chapters focuses on the construction of antibody libraries, antibody expression, complementary approaches for antibody selection, and other phage display related applications. 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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Phage Display: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
Table of Contents
Antibody phage display.- Construction of human immune and naive scFv Phage Display libraries.- Construction of Naïve and Immune Human Fab Phage Display Library.- Construction of synthetic antibody phage display libraries.- Construction of chicken and ostrich antibody libraries.- Construction of Rabbit Immune Antibody Libraries.- Isolation and characterization of single-domain antibodies from immune phage display libraries.- Phagekines: directed evolution and characterization of functional cytokines displayed on phages.- Efficient Cloning of Inserts for Phage Display by Golden Gate Assembly.- Construction of an ultra-large phage display library by Kunkel mutagenesis and rolling circle amplification.- Construction of semi-synthetic shark vNAR yeast surface display antibody libraries.- Antibody selection via Phage Display in microtiter plates.- Antibody selection in solution using magnetic beads.- Streptavidin Coated Solid Phase Extraction (SPE) TipsFor Antibody Phage Display Biopanning.- Magnetic Nanoparticle-Based Semi-Automated Panning for High-throughput Antibody Selection.- Antibody selection on cells targeting membrane proteinsTargeting Intracellular Antigens With pMHC-binding Antibodies: A Phage Display Approach.- Antibody Isolation from a Human Synthetic Libraries of Single-chain Antibodies and analysis using NGS.- Selection of affibody affinity proteins from phagemid libraries.- Antibody affinity and stability maturation by error-prone PCR.- Antibody batch cloning.- Deep Mining of Complex Antibody Phage Pools.- High-throughput IgG reformatting and expression using hybrid secretion signals and InTag positive selection technology.- Validation and the Determination of Antibody Bioactivity Using MILKSHAKE and Sundae Protocols.- Mapping polyclonal antibody responses to infection using next generation phage display.- Applications of high-throughput DNA sequencing to single-domain antibodydiscovery and engineering.- Biomarker discovery by ORFeome Phage Display.- Mapping Epitopes by Phage Display.- Epivolve: A Protocol for Site-directed Antibodies.
