Full Description
Since the advent of hybridoma technology more than two decades ago, numerous antibodies have entered the clinical setting as potent therapeutic agents. Their repeated application in humans, however, is limited by the development of human antimouse antibodies (HAMA) in the recipient, leading to allergic re- tions against the foreign murine protein and rapid neutralization. To circumvent these limitations many new antibodies have recently been tailored through recombinant antibody technology. The initial clinical data show encouraging results, thus demonstrating the potential of these new therapeutic agents. The purpose of Recombinant Antibodies for Cancer Therapy is to present a collection of detailed protocols in recombinant antibody technology. It is pri- rily addressed to scientists working on recombinant antibodies as well as cli- cians involved with antibody-based therapies. As with other volumes of this series, we placed the main focus on providing detailed protocols describing procedures step-by-step. Moreover, each protocol supplies a troubleshooting guide containing detailed information on possible problems and hints for pot- tial solutions. Antibody technology is a subject of constant and rapid change. This volume, therefore, does not attempt to cover all possible current experimental approaches in the field. Rather, we present carefully selected protocols, written by competent authors who have successfully verified the particular method described. Given our own professional backgrounds and interest in oncology, we chose to conc- trate chiefly on therapeutic agents for cancer patients.
Contents
Generation of Antibody Molecules Through Antibody Engineering.- Application of Recombinant Antibodies in Cancer Patients.- Hybridoma-Derived Antibodies.- DNA Immunization as a Means to Generate Antibodies to Proteins.- Chimerization of a Monoclonal Antibody for Treating Hodgkin's Lymphoma.- Humanization of Monoclonal Antibodies by CDR Grafting.- Recombinant Antibody Fragments from Phagemid-Displayed Antibody Repertoires.- Generation and Screening of a Modular Human scFv Expression Library from Multiple Donors.- Construction of Semisynthetic Antibody Libraries.- Single-Domain VH Antibody Fragments from a Phage Display Library.- Isolation of Human Fab Fragments Against Ovarian Carcinoma Using Guided Selection.- Human Recombinant Fab Antibodies with T-Cell Receptor-Like Specificities Generated from Phage Display Libraries.- Engineering Hot Spots for Affinity Enhancement of Antibodies.- Simultaneous Humanization and Affinity Optimization of Monoclonal Antibodies.- Tailoring Kinetics of Antibodies Using Focused Combinatorial Libraries.- Antibody Fragments with Additional Properties.- Engineering scFvs for Improved Stability.- Recombinant Single-Chain and Disulfide-Stabilized Fv Immunotoxins for Cancer Therapy.- Generation of Recombinant Immunotoxins for Specific Targeting of Tumor-Related Peptides Presented by MHC Molecules.- Construction and Characterization of RNase-Based Targeted Therapeutics.- Bispecific Diabodies for Cancer Therapy.- Generation and Characterization of Bispecific Tandem Diabodies for Tumor Therapy.- Generation of Recombinant Multimeric Antibody Fragments for Tumor Diagnosis and Therapy.- Construction and Characterization of Minibodies for Imaging and Therapy of Colorectal Carcinomas.- Generation, Expression, and Monitoring of Recombinant Immune Receptorsfor Use in Cellular Immunotherapy.- Tailoring Natural Effector Functions.- Single-Chain Fv-Based Affinity Purification of the Cellular Stress Protein gp96 for Vaccine Development.- Recombinant Adenoviruses for In Vivo Expression of Antibody Fragments.- Large Scale Production of Recombinant Antibodies for Clinical Application.- Production of Antibody Fragments in a Bioreactor.- Large Scale Production of Recombinant Antibodies by Utilizing Cellulose-Binding Domains.- Production of Tumor-Specific Antibodies in Tobacco.
