Soon after Kohler and Milstein described the use of somatic cell hybridization for the production of murine monoclonal antibodies of desired specificity, this relatively simple technique became widely applied. Indeed, production of murine monoclonal antibodies is now considered routine by immunologists and nonimmunologists alike. However, as heterologous proteins, mouse monoclonal antibodies have one major limitation: they are immunogenic in man and, hence, their use in vivo is severely limited. An obvious solution to this problem is to produce human hybridomas with the same techniques used…mehr
Soon after Kohler and Milstein described the use of somatic cell hybridization for the production of murine monoclonal antibodies of desired specificity, this relatively simple technique became widely applied. Indeed, production of murine monoclonal antibodies is now considered routine by immunologists and nonimmunologists alike. However, as heterologous proteins, mouse monoclonal antibodies have one major limitation: they are immunogenic in man and, hence, their use in vivo is severely limited. An obvious solution to this problem is to produce human hybridomas with the same techniques used for the production of rodent hybrids. Unfortunately, the history of human hybridomas has been marked by substantive and often exasperating tech nical problems, and the first reports of hybrids secreting human immu noglobulin of desired specificity did not appear until 1980. These reports were met with initial enthusiasm, but it soon became apparent that while human lymphocytes might be fused, their frequency, level of Ig synthesis, and stability were such that production of human antibodies with this method was neither routine nor practical. Nonetheless, a sufficient number of investiga tors persevered, and during the next 5 years relatively efficient B-cell fusion partners as well as improved methods of Epstein-Barr virus transformation were developed. Generation of human T -T hybrids has also been achieved, although problems of chromosomal stability remain a substantial obstacle, more so than with B-cell lines.Hinweis: Dieser Artikel kann nur an eine deutsche Lieferadresse ausgeliefert werden.
I B-Cell Lines, Hybridomas, and Monoclonal Antibodies.- A. Background and General Strategies.- 1 Human Hybridomas and Monoclonal Antibodies: The Biology of Cell Fusion.- 2 Fusion Partners for Production of Human Monoclonal Antibodies.- 3 Production of Human Monoclonal Antibodies Using Epstein-Barr Virus.- 4 The Epstein-Barr Virus-Hybridoma Technique.- 5 Strategies for Stable Human Monoclonal Antibody Production: Construction of Heteromyelomas, in Vitro Sensitization, and Molecular Cloning of Human Immunoglobin Genes.- B. Applications to Infectious Diseases.- 6 Production and Characterization of Human Monoclonal Antibodies against Gram-Negative Bacteria.- 7 Human Monoclonal Antibodies to Defined Antigens: Toward Clinical Applications.- 8 Production of Human Monoclonal Antibodies Using a Human-Mouse Fusion Partner.- 9 In Vitro Expansion of Human B Cells for the Production of Human Monoclonal Antibodies.- 10 Cell-Driven Viral Transformation.- C. Applications to Cancer.- 11 The Generation of Human Monoclonal Antibodies and Their Use in the Analysis of the Humoral Immune Response to Cancer.- 12 Design and Production of Human Monoclonal Antibodies to Human Cancers.- 13 Human-Human Hybridoma Technology: Five Years of Technical Improvements, and Its Application in Cancer Biology.- D. Applications to Autoimmunity.- 14 The Production of Monoclonal Antibodies by Human-Human Hybridomas: Their Application to Studies of Autoimmune Diseases.- 15 Human Monoclonal Autoantibodies Reactive with Multiple Organs.- 16 Principles of in Vitro Immunization of Human B Lymphocytes.- E. Special Topics.- 17 Human-Human Hybridomas in the Study of Immunodeficiencies.- II Human T-T Hybridomas.- 18 Human T-Lymphocyte Subsets and T-T Hybridomas: An Overview.- 19 Production of Human T-Cell Hybridomasby Electrofusion.- 20 Human B-Cell Growth and Differentiation Factors Secreted by T-Cell Hybridomas.- 21 Generation and Characterization of Human T-Cell Hybridomas That Constitutively Produce Immune Interferon.- 22 Selection of Human T-Cell Hybridomas That Produce Inflammatory Lymphokines by the Emetine-Actinomycin D Method.- 23 Human T-T Hybridomas Specific for Epstein-Barr Virus: Generation and Function.- 24 Factors Generated by Human T-Cell Hybridomas Regulate B-Cell Activation, Polyclonal Differentiation, and Isotype Expression.- 1. Human T- and B-Cell Lines.- 2. Preparation of Mutant Cell Lines.- 3. Freezing Human Cell Lines.- 4. Mycoplasma Testing.- 5. Peripheral Blood Lymphocyte Separation from Whole Blood or Buffy Coats.- 6. Separation of Human T and Non-T Lymphocytes from Peripheral Blood.- 7. Panning for Human T-Lymphocyte Subpopulations.- 8. Human Hybridoma Tube Fusion Protocol.- 9. Plate Fusion Technique for Nonadherent Cells.- 10. Identification of Human Chromosomes in Mouse-Human Lymphocyte Hybrids.- 11. Epstein-Barr Virus Transformation.- 12. Growth of Human Cell Lines in Mice.- 13. Methods of Large-Scale Tissue Culture.- 16. Soft Agar Cloning Protocol.- 17. Cloning by Limiting Dilution.- 18. Reverse Plaque Assay.- 19. Assay of Interleukin 1 (IL-1).- 20. Assay of Interleukin 2 (IL-2).- 21. Assay of B-Cell Growth and Differentiation Factors.- 22. Indirect Immunoglobulin ELISA Protocol.- 23. Purification of Human Immunoglobulins.- 24. A Nitrocellulose Strip Method for Isotyping Monoclonal Antibodies.- 25. Immunoprecipitation of Antigens Using Polystyrene Balls.- 26. Immunostaining Using Monoclonal Antibodies.- 27. Immunoblotting.- Manufacturers and Distributors.
I B-Cell Lines, Hybridomas, and Monoclonal Antibodies.- A. Background and General Strategies.- 1 Human Hybridomas and Monoclonal Antibodies: The Biology of Cell Fusion.- 2 Fusion Partners for Production of Human Monoclonal Antibodies.- 3 Production of Human Monoclonal Antibodies Using Epstein-Barr Virus.- 4 The Epstein-Barr Virus-Hybridoma Technique.- 5 Strategies for Stable Human Monoclonal Antibody Production: Construction of Heteromyelomas, in Vitro Sensitization, and Molecular Cloning of Human Immunoglobin Genes.- B. Applications to Infectious Diseases.- 6 Production and Characterization of Human Monoclonal Antibodies against Gram-Negative Bacteria.- 7 Human Monoclonal Antibodies to Defined Antigens: Toward Clinical Applications.- 8 Production of Human Monoclonal Antibodies Using a Human-Mouse Fusion Partner.- 9 In Vitro Expansion of Human B Cells for the Production of Human Monoclonal Antibodies.- 10 Cell-Driven Viral Transformation.- C. Applications to Cancer.- 11 The Generation of Human Monoclonal Antibodies and Their Use in the Analysis of the Humoral Immune Response to Cancer.- 12 Design and Production of Human Monoclonal Antibodies to Human Cancers.- 13 Human-Human Hybridoma Technology: Five Years of Technical Improvements, and Its Application in Cancer Biology.- D. Applications to Autoimmunity.- 14 The Production of Monoclonal Antibodies by Human-Human Hybridomas: Their Application to Studies of Autoimmune Diseases.- 15 Human Monoclonal Autoantibodies Reactive with Multiple Organs.- 16 Principles of in Vitro Immunization of Human B Lymphocytes.- E. Special Topics.- 17 Human-Human Hybridomas in the Study of Immunodeficiencies.- II Human T-T Hybridomas.- 18 Human T-Lymphocyte Subsets and T-T Hybridomas: An Overview.- 19 Production of Human T-Cell Hybridomasby Electrofusion.- 20 Human B-Cell Growth and Differentiation Factors Secreted by T-Cell Hybridomas.- 21 Generation and Characterization of Human T-Cell Hybridomas That Constitutively Produce Immune Interferon.- 22 Selection of Human T-Cell Hybridomas That Produce Inflammatory Lymphokines by the Emetine-Actinomycin D Method.- 23 Human T-T Hybridomas Specific for Epstein-Barr Virus: Generation and Function.- 24 Factors Generated by Human T-Cell Hybridomas Regulate B-Cell Activation, Polyclonal Differentiation, and Isotype Expression.- 1. Human T- and B-Cell Lines.- 2. Preparation of Mutant Cell Lines.- 3. Freezing Human Cell Lines.- 4. Mycoplasma Testing.- 5. Peripheral Blood Lymphocyte Separation from Whole Blood or Buffy Coats.- 6. Separation of Human T and Non-T Lymphocytes from Peripheral Blood.- 7. Panning for Human T-Lymphocyte Subpopulations.- 8. Human Hybridoma Tube Fusion Protocol.- 9. Plate Fusion Technique for Nonadherent Cells.- 10. Identification of Human Chromosomes in Mouse-Human Lymphocyte Hybrids.- 11. Epstein-Barr Virus Transformation.- 12. Growth of Human Cell Lines in Mice.- 13. Methods of Large-Scale Tissue Culture.- 16. Soft Agar Cloning Protocol.- 17. Cloning by Limiting Dilution.- 18. Reverse Plaque Assay.- 19. Assay of Interleukin 1 (IL-1).- 20. Assay of Interleukin 2 (IL-2).- 21. Assay of B-Cell Growth and Differentiation Factors.- 22. Indirect Immunoglobulin ELISA Protocol.- 23. Purification of Human Immunoglobulins.- 24. A Nitrocellulose Strip Method for Isotyping Monoclonal Antibodies.- 25. Immunoprecipitation of Antigens Using Polystyrene Balls.- 26. Immunostaining Using Monoclonal Antibodies.- 27. Immunoblotting.- Manufacturers and Distributors.
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