Note: Front Cover -- Monoclonal Antibody Technology -- Copyright Page -- Contents -- Preface -- Acknowledgements -- Abbreviations -- Chapter 1. General properties and applications of monoclonal antibodies -- 1.1. Introduction -- 1.2. Comparison of monoclonal antibodies and conventional antiserum -- 1.3. Application of monoclonal antibodies -- Chapter 2. Assay techniques -- 2.1. General assay requirements -- 2.2. Theoretical considerations -- 2.3. Practical considerations -- 2.4. Antibody sampling -- 2.5. Types of assay -- 2.6. Solid-phase-assays -- 2.7. Soluble-phase systems -- 2.8. Cellular assays -- 2.9. Biological assays -- 2.10 Immunocytochemical assays -- Chapter 3. Selection of animals and cell lines -- 3.1. Choice of animal -- 3.2. Choice of cell lines -- 3.3. Selective drug markers -- 3.4. The mouse system -- 3.5. The rat system -- 3.6. The human system -- 3.7. T cell lines -- Chapter 4. Immunisation -- 4.1. Introduction -- 4.2. Immunisation schedules -- 4.3. Immunisation in vitro -- 4.4. Isotype enrichment -- Chapter 5. Cell culture requirements for hybridomas -- 5.1. Introduction -- 5.2. Basic requirements -- 5.3. Basic cell-culture techniques -- 5.4. Contamination -- 5.5. Feeder cells -- Chapter 6. Fusion procedures -- 6.1. The use of polyethylene glycol -- 6.2. The components of HAT medium -- 6.3. Selection of hybridomas by fluorescence activated cell sorting -- 6.4. Preparation of stock solutions -- 6.5. Fusion frequencies and plating densities -- 6.6. Fusion protocols for mouse experiments -- Chupter 7. Transformation -- 7.1. Introduction -- 7.2. Pre-selection of lymphocytes of pre-defined specificity -- 7.3. Pre-selection of T lymphocytes -- 7.4. Transformation -- 7.5. Transformation followed by fusion -- Chapter 8. Selection and cloning -- 8.1. Early feeding and assay of fusions -- 8.2. Failure of fusions -- 8.3. Cloning of hybridomas. , 8.4. Failure of cloning -- 8.5. Continuation of cloning -- Chapter 9. Antibody production and purification -- 9.1. Maintenance of cell stocks -- 9.2. Expansion of hybridomas in vitro -- 9.3. Expansion of hybridomas in vivo -- 9.4. Failure in expansion -- 9.5. Storage of antibody -- 9.6. Concentrating the antibody -- 9.7. Purification of the antibody -- Chapter 10. Characterisation of monoclonal antibodies -- 10.1. Introduction -- 10.2. Determination of antibody class -- 10.3. Proof that the antibody is monoclonal -- 10.4. Epitope analysis -- 10.5. Determination of antibody affinity -- 10.6. Karyotype analysis of hybridomas -- Appendix 1. Animal handling techniques -- Appendix 2. Addresses of suppliers and manufacturers -- Appendix 3. Protocols for polyacrylamide gel electrophoresis (PAGE) -- References -- Addendum -- Subject Index.

Note: Front Cover -- Monoclonal Antibody and Immunosensor Technology -- Copyright Page -- Contents -- Preface -- Acknowledgements -- Abbrevations -- Chapter 1. General properties and applications of monoclonal antibodies -- 1.1. History of Mab technology -- 1.2. Introduction to Mab technology -- 1.3. Specificity/affinity - the contribution of monoclonal antibody -- 1.4. Specificity/affinity - the contribution of antigen -- 1.5. Specificity/affinity - the contribution of the assay system -- 1.6. Pooling of Mabs -- 1.7. The standard nature of Mabs -- 1.8. Yield and purity -- 1.9. Human monoclonal antibodies (HuMabs) -- 1.10. Chimaeric bifunctional monoclonal antibodies -- 1.11. Applications of Mabs - Diagnostic -- 1.12. Therapeutic applications -- 1.13. Mabs in vaccine development strategies -- 1.14. Mabs as catalysts (abzymes) -- 1.15. Mabs in purification -- 1.16. Mabs in basic immunological research -- 1.17. Patent considerations for Mabs -- 1.18. Buying a Mab -- Chapter 2. Assay techniques -- 2.1. General assay requirements -- 2.2. Theoretical considerations -- 2.3. Practical screening in the lab -- 2.4. Antibody sampling -- 2.5. Types of assay -- 2.6. Solid-phase assays -- 2.7. Protocols for solid-phase assays -- 2.8. Screening for cellular antigens by FACS -- 2.9. Screening for cellular antigens by immunocytochemical methods -- 2.10. Screening for catalytic antibodies -- 2.11. Screening recombinant libraries -- Chapter 3. Selection of animals, tissues and cell lines -- 3.1. Animals in which hybridomas may be generated -- 3.2. Whether to choose the rat or mouse system -- 3.3. Selective drug markers -- 3.4. The mouse system -- 3.5. The rat system -- 3.6. The human system -- 3.7. T cell lines -- 3.8. Tissues used as sources of B cells -- Chapter 4. Immunisation -- 4.1. How the natural immune response operates -- 4.2. Is it necessary to immunise?. , 4.3. Is it necessary to know precisely what the antigen is? -- 4.4. Purity of antigen -- 4.5. The use of previously determined schedules -- 4.6. Strains of rodent -- 4.7. Age and sex of rodent -- 4.8. Tolerance, overimmunisation and underimmunisation -- 4.9. Adjuvants -- 4.10. In vitro immunisation -- 4.11. Production of specific antibody isotypes -- 4.12. Routes of immunisation -- 4.13. Schedules -- 4.14. Specific examples - proteins -- 4.15. Specific examples - carbohydrates -- 4.16. Specific examples - lipids and nucleic acids -- 4.17. Specific examples - haptens -- 4.18. Specific examples - whole cells and organisms -- 4.19. Immunisation of human subjects -- Chapter 5. Cell culture requirements for hybridomas -- 5.1. Introduction -- 5.2. Basic cell culture requirements -- 5.3. Basic cell culture techniques -- 5.4. Contamination -- 5.5. Feeder cells -- Chapter 6. Fusion procedures -- 6.1. The use of polyethylene glycol -- 6.2. The components of HAT medium -- 6.3. Selection of hybridomas by fluorescent activated cell sorting (FACS) -- 6.4. Preparation of stock solutions -- 6.5. Fusion frequencies and plating densities -- 6.6. Fusion protocols for mouse experiments -- 6.7. Electrofusion -- Chapter 7. Alternative strategies for the immortalisation of antibody producing cells -- 7.1. Introduction -- 7.2. Pre-selection of lymphocytes with defined cell surface markers or antigen binding capacity -- 7.3. Pre-selection of T lymphocytes -- 7.4. Transformation of B lymphocytes -- 7.5. Post-transformation strategies -- 7.6. Transformation with oncogenes -- 7.7. Short term cloning of B cells in lymphokine enriched medium -- Chapter 8. Selection and cloning -- 8.1. Early feeding and assay of fusions -- 8.2. Failure of fusions -- 8.3. Cloning of hybridomas -- 8.4. Failure of cloning -- 8.5. Continuation of cloning. , Chapter 9. The use of recombinant DNA methods -- 9.1. Background -- 9.2. Antibody genes -- 9.3. Whether to make whole antibody, or its subfragments -- 9.4. The steps involved in cloning of antibody genes -- 9.5. Expression in E. coli -- 9.6. Recombinant techniques involving the use of PCR -- 9.7. Where recombinant DNA technology will be of value -- Chapter 10. Antibody production and purification -- 10.1. Maintenance of cell stocks -- 10.2. Expansion of hybridomas on a laboratory scale (1-20 mg Ab) -- 10.3. Expansion in vitro for more extensive characterisation -- 10.4. Expansion of hybridomas in vivo -- 10.5. Storage of Mabs -- 10.6. Purification of Mabs -- 10.7. Purification of IgM Mabs -- 10.8. Purification of IgA Mabs -- Chapter 11. Characterisation of monoclonal antibodies -- 11.1. Introduction -- 11.2. Determination of antibody class -- 11.3. Proof that the antibody is monoclonal -- 11.4. Epitope analysis -- 11.5. Characterisation of cell surface antigens -- 11.6. Determination of antibody affinity -- 11.7. Karyotype analysis of hybridomas -- Chapter 12. Immunosensors -- 12.1. Biosensors -- 12.2. Enzyme biosensors using enzymes as sensing elements -- 12.3. The difference between enzyme based sensing elements and antibody based sensing elements -- 12.4. Requirement and potential application of antibody based immunosensors -- 12.5. Classification of immunosensor systems -- 12.6. General rules for adapting a Mab or panel of Mabs into a biosensor system -- 12.7. Labelling Mabs and their antigens -- Appendix 1 - Animal handling techniques -- A.1.1. Introduction -- A.1.2. Requirement for animal handling licence -- A.1.3. Handling -- A.1.4. Immunisation -- A.1.5. Serum collection -- A.1.6. Removal of tissues at sacrifice -- A.1.7. Ascites fluid -- Appendix 2 - Addresses of Suppliers and Manufacturers. , Appendix 3 - Protocols for polyacrylamide gel electrophoresis (PAGE) -- A.3.1. Introduction -- A.3.2. Slab gel apparatus -- A.3.3. SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) -- A.3.4. Isoelectric focussing of immunoglobulins -- A.3.5. Electrophoresis of DNA on agarose -- A.3.6. Autoradiography or fluorography of gels -- References -- Subject index.