Lo Benny K.C. (Ed.) Antibody Engineering: Methods and Protocols

Monograph. Antibody engineering methods and protocols.
Springer, 2004 - 561 p.A core collection of diverse cutting-edge techniques for the generation, expression, optimization, and characterization of recombinant antibodies. Readily reproducible protocols for lead generation range from the cloning of human immunoglobulin genes to the selection and generation of human recombinant antibodies by humanization approaches, molecular display technologies and transgenic animals. Procedures are also described on restructuring antibody leads into monovalent, multivalent, and bispecific binding fragments for a wide variety of in vivo applications. State-of-the-art technologies are described for the characterization of antigen-binding affinity and specificity with novel applications in radioimmunotargeting, cancer immunotherapy, drug abuse, and proteomics. Use cutting-edge techniques for generating and optimizing recombinant antibodies Generate antibodies by humanization, molecular display technologies and transgenic mice Perform epitope mapping with mass spectrometry, bioinformatics, and array technologies Learn about novel applications in cancer, drug addiction, and proteomics Express recombinant antibodies in bacterial, yeast, insect, mammalian and plant systems.Internet Resources for the Antibody Engineer.
The Kabat Database and a Bioinformatics Example.
IMGT, The International ImMunoGeneTics Information System.
Antibody Variable Regions: Toward a Unified Modeling Method.
Studying Antibody Conformations by Ultracentrifugation and Hydrodynamic Modeling.
PCR Cloning of Human Immunoglobulin Genes.
Antibody Humanization by CDR Grafting.
Selection of Human Antibodies from Phage Display Libraries.
Production of Human Single-Chain Antibodies by Ribosome Display.
Production of Human Antibodies from Transgenic Mice.
Selection of Internalizing Antibodies for Drug Delivery.
Engineering Multivalent Antibody Fragments for In Vivo Targeting.
Production of Recombinant Bispecific Antibodies.
Expression and Isolation of Recombinant Antibody Fragments in E. coli.
Expression of Recombinant Antibodies in Mammalian Cell Lines.
Human Antibody Production Using Insect-Cell Expression Systems.
Antibody Production in Transgenic Plants.
Directed Mutagenesis of Antibody Variable Domains.
Antibody Affinity Maturation by Chain Shuffling.
Antibody Affinity Maturation by Random Mutagenesis.
Developing a Minimally Immunogenic Humanized Antibody by SDR Grafting.
Antibody Purification by Column Chromatography.
Affinity Measurement Using Surface Plasmon Resonance.
Kinetic Exclusion Assays to Study High-Affinity Binding Interactions in Homogeneous Solutions.
Characterization of Antibody-Antigen Interactions by Fluorescence Spectroscopy.
Antibody Epitope Mapping Using Arrays of Synthetic Peptides.
Mapping Antibody:Antigen Interactions by Mass Spectrometry and Bioinformatics.
Radiometal Labeling of Antibodies and Antibody Fragments for Imaging and Therapy.
Production and Characterization of Anti-Cocaine Catalytic Antibodies.
Recombinant Immunotoxins in the Treatment of Cancer.
Antibodies in Proteomics.
Targeting of Antibodies Using Aptamers.