Table of Contents
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.
