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.With 10 recent FDA approvals of therapeutic antibody drugs and hundreds more in the biotechnology pipeline, it is clear that engineered antibodies will play a major role in treating human diseases in the 21st century. In Antibody Engineering: Methods and Protocols, leading researchers and clinicians present a core collection of 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 generation of human recombinant antibodies by humanization approaches, molecular display technologies, and transgenic animals. The techniques for antibody expression and optimization utilize not only bacterial and mammalian cell cultures, but also insect cell cultures and transgenic plants. 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. Each fully tested protocol is described in step-by-step detail and includes background material, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls.§Comprehensive and up-to-date, Antibody Engineering: Methods and Protocols provides both laboratory and clinical investigators with a diverse array of the fundamental techniques upon which new and imaginative drug technologies can be successfully developed.
