Monoclonal Antibodies The Second Generation

Contributors -- Abbreviations -- Preface -- Safety note -- 1. Introduction. H. Zola -- Introduction: the purpose of this book -- Monoclonal antibodies: scope and limitations -- Antibodies as laboratory reagents: a brief perspective -- Antibodies as therapeutic agents: a brief perspective -- Conclusions -- References -- 2. Making and using 'conventional' mouse monoclonal antibodies. P.J. Donohoe, P.J. Macardle and H. Zola -- Introduction -- Tissue culture -- Materials -- Working solutions -- The screening assay -- Immunization -- The myeloma cell line -- Fusion protocol -- Feeder cells -- Post-fusion care -- Mycoplasma -- Cloning -- Cryopreservation -- Thawing cells out -- Ascites fluid -- Production of monoclonal antibodies in dialysis tubing -- Continuous perfusion culture systems -- Purification -- Conjugation -- References -- 3. Human monoclonal antibodies from immortalized B cells. A. W. Hohmann, L. Spatz, M. Irigoyen and A. Manheimer-Lory -- Introduction -- Immortalized B cells to probe the human antibody repertoire -- Infectious diseases -- Human immunodeficiency virus -- Erythrocyte antigens -- Human histocompatibility antigens -- Tumor antigens -- Autoantibodies -- Immortalization of B cells with EBV -- EBV transformation of B cells: methodology -- Making human hybridomas -- Hybridoma methodology -- References -- 4. Reconstruction of monoclonal antibodies by genetic engineering. J.R. Adair and D.J. King -- Introduction -- The reconstruction process -- The binding site -- The effector functions -- Reconstructed antibodies in practice -- Summary and conclusions -- References -- 5. The genetic engineering of antibody constructs for diagnosis and therapy. G.A. Pietersz and l.F.C. McKenzie -- Introduction -- Principles of use of antibody conjugates -- Recent advances in immunoconjugate construction -- Recombinant antibodies and fusion proteins -- Use of cytokines and growth factors for targeting -- Potential problems -- Conclusions -- References -- 6. Superseding hybridoma technology with phage display libraries. R. Irving and P. Hudson -- Introduction -- Affinity maturation -- Discussion -- References -- 7. Construction and application of libraries of artificial antibodies. W. Ito and Y. Kurosawa -- Introduction -- Expression in E. coli of antibodies whose V genes are derived from V" and V L genes expressed in animals -- Characteristics of phage display antibodies -- Various devices for the generation of artificial antibodies -- Construction of libraries of completely synthetic antibodies -- Application of libraries of artificial antibodies -- Future perspectives -- References -- 8. Expression of antibody genes in mammalian cells. C. Bebbington -- Introduction -- Transient expression -- Stable mammalian expression systems -- Other factors influencing expression in mammalian cells -- Choosing an expression system -- References -- 9. Structure and application of single-chain Fvs as diagnostic and therapeutic agents. P. Hudson -- Introduction -- Design of single-chain Fv fragments -- Construction of scFv genes -- The structure of scFv fragments -- The design of fusion proteins -- The design of bifunctional scFvs ( diabodies) -- The choice of expression systems -- Scale-up production of scFvs in fermenters -- Case studies: the design and expression of scFv molecules (for crystal structures and novel diagnostics) -- Applications of scFvs in diagnostics -- Discussion -- References -- Index.

Biography

Chief Hospital Scientist and Professor of Immunology, Flinders Medical Centre and Flinders University, Adelaide, Australia.