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.

InhaltsangabePart I. Introduction Generation of Antibody Molecules Through Antibody Engineering Sergey M. Kipriyanov Application of Recombinant Antibodies in Cancer Patients Jürgen Krauss, Michaela Arndt, and Michael Pfreundschuh Part II. Hybridoma-Derived Antibodies DNA Immunization as a Means to Generate Antibodies to Proteins Partha S. Chowdhury Chimerization of a Monoclonal Antibody for Treating Hodgkin's Lymphoma Jürgen Krauss, Hans Heinrich Förster, Barbara Uchanska-Ziegler, and Andreas Ziegler Humanization of Monoclonal Antibodies by CDR Grafting Siobhan O'Brien and Tarran Jones Part III. Recombinant Antibody Fragments from Phagemid-Displayed Antibody Repertoires Generation and Screening of a Modular Human scFv Expression Library from Mulitple Donors Martin Welschof, Christiane Christ, Ingrid Hermes, Armin Keller, Christian Kleist, and Michael Braunagel Construction of Semisynthetic Antibody Libraries Michael Braunagel SingleDomain VH Antibody Fragments from a Phage Display Library Liat Binyamin, Daniel Plaksin, and Yoram Reiter Isolation of Human Fab Fragments Against Ovarian Carcinoma Using Guided Selection Mariangela Figini, Andrew Green, Francesco Colotta, and Silvana Canevari Human Recombinant Fab Antibodies with T-Cell Receptor-Like Specificities Generated from Phage Display Libraries Jan Engberg, Ali F. Yenidunya, Rikke Clausen, Liselotte B. Jensen, Peter Sørensen, Pernille Kops, and Erik Riise Engineering Hot Spots for Affinity Enhancement of Antibodies Partha S. Chowdhury Simultaneous Humanization and Affinity Optimization of Monoclonal Antibodies Herren Wu Tailoring Kinetics of Antibodies Using Focused Combinatorial Libraries Herren Wu and Ling-Ling An Part IV. Antibody Fragments with Additional Properties Engineering scFvs for Improved Stability Partha S. Chowdhury and George Vasmatzis Recombinant Single-Chain and Disulfide-Stabilized Fv Immunotoxins for Cancer Therapy Revital Niv, Dina Segal, and Yoram Reiter Generation of Recombinant Immunotoxins for Specific Targeting of Tumor-Related Peptides Presented by MHC Molecules Cyril J. Cohen, Galit Denkberg, Dina Segal, and Yoram Reiter Construction and Characterization of RNase-Based Targeted Therapeutics Dianne L. Newton, Junichiro Futami, Dale Ruby, and Susanna M. Rybak Bispecific Diabodies for Cancer Therapy Michaela Arndt and Jürgen Krauss Generation and Characterization of Bispecific Tandem Diabodies for Tumor Therapy Sergey M. Kipriyanov Generation of Recombinant Multimeric Antibody Fragments for Tumor Diagnosis and Therapy Barbara E. Power, Alexander A. Kortt, and Peter J. Hudson Construction and Characterization of Minibodies for Imaging and Therapy of Colorectal Carcinomas Paul J. Yazaki and Anna M. Wu Generation, Expression, and Monitoring of Recombinant Immune Receptors for Use in Cellular Immunotherapy Andreas Hombach, Claudia Heuser, and Hinrich Abken Tailoring Natural Effector Functions: Antibody Engineering Beyond Humanization Ole H. Brekke and John E. Thommesen SingleChain FvBased Affinity Purification of the Cellular Stress Protein gp96 for Vaccine Development Christian Kleist, Daniele Arnold-Schild, Martin Welschof, Martina Finger, Gerhard Opelz, Hans-Georg Rammensee, Hansjörg Schild, and Peter Terness Recombinant Adenoviruses for In Vivo Expression of Antibody Fragments Roland E. Kontermann, Tina Korn, and Valérie Jérôme Part V. Large Scale Production of Recombinant Antibodies for Clinical Application Production of Antibody Fragments in a Bioreactor Halldis Hellebust. Large Scale Production of Recombinant Antibodies by Utilizing Cellulose-Binding Domains Itai Benhar and Yevgeny Berdichevsky Production of Tumor-Specific Antibodies in Tobacco Carmen Vaq

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