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

Монография. Инженерия антител методы и протоколы
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, http://imgt.cines.fr
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