Antibody phage display, the definitive technology for monoclonal production, has now advanced to a stage where it can be performed in nonspecialized research laboratories. In Antibody Phage Display: Methods and Protocols, Philippa M. O'Brien and Robert Aitken combine in one volume a comprehensive collection of established antibody phage display protocols, each accompanied by authoritative guidance that will enable the nonspecialist to carry them out successfully. Coverage spans the construction of antibody libraries, the selection of antibody clones with the desired properties, and their modification, expression, and purification. Each readily reproducible method is described by a hands-on expert in step-by-step detail and includes a wealth of practical advice not found in the scientific literature. Extensive notes discuss pitfalls to avoid and offer many possible alternative methods to suit special research situations. An overview by one of the world's leading experts in antibody phage display, Dr. Hennie R. Hoogenboom, surveys the current status of the field and the future of the technology.Comprehensive and highly practical, Antibody Phage Display: Methods and Protocols provides biochemists, molecular biologists, and immunologists with a gold-standard reference guide to the successful isolation, modification, and expression of recombinant antibodies using today's powerful phage display technology.

Overview of Antibody Phage-Display Technology and Its Applications.- Standard Protocols for the Construction of Fab Libraries.- Standard Protocols for the Construction of scFv Libraries.- Broadening the Impact of Antibody Phage Display Technology.- Construction of Large Naïve Fab Libraries.- Construction of Polyclonal Antibody Libraries Using Phage Display.- Antigen-Driven Stimulation of B-Lymphocytes In Vitro.- 8 The Recovery of Immunoglobulin Sequences from Single Human B Cells by Clonal Expansion.- Panning of Antibody Phage-Display Libraries.- Selection of Antibodies Against Biotinylated Antigens.- Isolation of Anti-Hapten Specific Antibody Fragments from Combinatorial Libraries.- Blocking Immunodominant Epitopes by Competitive Deselection.- Rescue of a Broader Range of Antibody Specificities Using an Epitope-Masking Strategy.- Screening of Phage-Expressed Antibody Libraries by Capture Lift.- Antibody-Guided Selection Using Capture-Sandwich ELISA.- Proximity-Guided (ProxiMol) Antibody Selection.- Isolation of Human Monoclonal Antibodies Using Guided Selection with Mouse Monoclonal Antibodies.- Selecting Antibodies to Cell-Surface Antigens Using Magnetic Sorting Techniques.- Isolation of Human Tumor-Associated Cell Surface Antigen-Binding scFvs.- Subtractive Isolation of Single-Chain Antibodies Using Tissue Fragments.- Selection of Antibodies Based on Antibody Kinetic Binding Properties.- Selection of Functional Antibodies on the Basis of Valency.- Two-Step Strategy for Alteration of Immunoglobulin Specificity by In Vitro Mutagenesis.- Targeting Random Mutations to Hotspots in Antibody Variable Domains for Affinity Improvement.- Error-Prone Polymerase Chain Reaction for Modification of scFvs.- Use of Escherichia coli Mutator Cells to Mature Antibodies.- ChainShuffling to Modify Properties of Recombinant Immunoglobulins.- Generation of Bispecific and Tandem Diabodies.- High-Level Periplasmic Expression and Purification of scFvs.- Periplasmic Expression and Purification of Recombinant Fabs.- Expression of Antibody Fragments in Pichia pastoris.- Expression of VHH Antibody Fragments in Saccharomyces cerevisiae.- Intrabodies.- 34 Expression of scFvs and scFv Fusion Proteins in Eukaryotic Cells.- 35 Expression of Antibody Fab Fragments and Whole Immunoglobulin in Mammalian Cells.