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.

Cloning of the Antigen-binding Site from Hybridoma

Cloning of Variable Domains from Mouse Hybridoma by PCR
Coning Hybridoma cDNA by RACE
Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly
Generation of Antibody Repertoires
Mouse Immune Libraries for the Generation of ScFv Fragments Directed Against Human Cell Surface Antigens
Human Antibody Gene Libraries
Synthetic Antibody Libraries
Immune Libraries from Nonhuman Primates (NHP)
Generation of Rabbit Immune Libraries
Selection of Antibody Fragments from Combinatorial Libraries
Immunotube Selections
Phage Display and Selection in Microtitre Plates
Phage Display and Selections on Biotinylated Antigens
Phage Display and Subtractive Selection on Cells
Selection of Phage Antibody Libraries for Binding and Internalization into Mammalian Cells
Improving Phage Display Throughput by Using Hyperphage, Miniaturized Titration and pVIII (g8p) ELISA
Yeast Display and Selections
The Generation of Transgenic Mice Expressing Human Antibody Repertoires
Selection of Antibody Fragments by Means of the Filter-Sandwich Colony Screening Assay
Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries
Engineering and Production of Immunoglobulins
Aspects of Isotype Selection
Generation of Heavy and Light Chains (Chimeric Antibodies)
Humanising Antibodies by CDR Grafting
Humanization by Resurfacing
Human Antibodies by Guided Selection
In Silico De-Immunization
Affinity Maturation by Chain Shuffling and Site Directed Mutagenesis
Affinity Maturation by Random Mutagenesis and Phage Display
Engineering of the Fc Region for Improved PK (FcRn Interaction)
Antibody-Dependent Enzyme Prodrug Therapy (ADEPT)
Production of Recombinant Human IgG Antibodies in the Baculovirus Expression System
Expression of IgA Molecules in Mammalian Cells
Antibody Characterization
Expression of Complete Antibodies in Transgenic Plants
Expression ofFull Length Monoclonal Antibodies (mAb) in Algal Chloroplast
Expression of IgG Antibodies in Mammalian Cells
Protein A/G Chromatography
Epitope Analysis Using Synthetic Peptide Repertoires Prepared by SPOT Synthesis Technology
Epitope Mapping by Printed Peptide Libraries
Antibody Epitope Mapping Using Yeast Display
Size Exclusion Chromatography
Structural Characterization of Antibodies by Mass Spectrometry
Antibody Glycans Characterization
Affinity Measurements by Competition ELISA
Anti-Histidine Antibodies as Tools for Reversible Capturing of His-Tagged Fusion Proteins for Subsequent Binding Analysis
Affinity Measurements Using Quartz Crystal Microbalance (QCM)
Affinity Measurements with Radiolabeled Antibodies
Neutralization Tests
Functional Characterization of Antibodies Neutralizing Soluble Factors In Vitro and In Vivo
Competitive ELISA
Quantification of Human IgG and Related Fc Fusion Proteins by a Human IgG/Fc Capture ELISA
Determination of Fc-Mediated Antibody-Effector Functions by Chromium Release Assay
Binding Studies with Flow Cytometry.