A sample of the most exciting developments in the cloning, manipulation, expression and application of genetically-engineered monoclonal antibodies. This rapidly-evolving field has witnessed the PCR combinatorial cloning of vast immunological diversity, in vitro mutagenesis of MAbs, MAbs created by transgenic animals, novel expression systems in plants, animals and lower systems, as well as a rich variety of genetically modified MAbs as potential therapeutic agents. Leading scientists from academia and industry present their own findings as well as short reviews of these research areas.
Inhalt
Section I: Human Monoclonal Antibodies.- 1 Human Monoclonal Antibody Technology.- A. Introduction.- I. Why Produce Human Monoclonals?.- II. Chapter Aims.- III. Approach Used.- B. General Production Strategies.- I. Introduction.- II. Source of Immune Lymphocytes.- 1. Usual Sources.- 2. In Vitro Immunization.- 3. Generating Immune Lymphocytes in Severe Combined Immunodeficiency Mice.- III. Processing of Lymphoid Tissues.- IV. Immortalization Strategies.- 1. Introduction.- 2. Cell Fusion.- 3. Epstein-Barr Virus Fusion.- 4. Combined Epstein-Barr Virus Transformation and Cell Fusion.- 5. Novel Approaches.- V. Selection, Cloning and Expansion.- 1. Introduction.- 2. Selection.- 3. Cloning.- 4. Expansion.- 5. Additional Evaluation.- VI. Bispecific and Trispecific Antibodies.- C. Human Monoclonal Targets.- I. Antibody Specificities Generated.- II. Application of Human Monoclonal Antibody Technology.- 1. Introduction.- 2. Tumour Field.- 3. AIDS Research.- 4. Autoimmunity.- 5. Future Targets.- D. Limitations of Orthodox Technology.- I. Introduction.- II. Why Is Antibody Secretion Unsatisfactory?.- III. Is Unsatisfactory Secretion Related to Cell Surface Phenotype or Cytokine Secretion?.- E. Conclusion.- I. Impact and Potential of Recombinant Technology.- 1. Chimeric and Humanized Antibodies.- 2. Repertoire Cloning.- 3. Combining Cell and Gene Cloning Technologies.- References.- 2 Recombinant Therapeutic Human Monoclonal Antibodies.- A. Therapeutic Human Monoclonal Antibodies.- B. Rapid Direct Cloning of Antibody Variable Regions.- C. Genetically Engineered Chimeric Monoclonal Antibodies.- I. Chimeric Antibodies.- 1. Summary of Work with Therapeutic Chimeric Monoclonal Antibodies.- II. Recombinant Conjugates and Fusion Proteins.- 1. Immunotoxins.- 2. A Recombinant Monoclonal Antibody Linked to Tissue-Type Plasminogen Activator.- 3. T Cell Receptor Conjugates.- 4. Growth Factor Conjugates.- 5. Other Fusion Proteins.- 6. Antibody-Enzyme Conjugates for Cancer.- D. Reshaped or Composite Antibodies.- E. Immortalization of the Immunoglobulin Repertoire Using rDNA Technology.- F. Recombinatorial Antibody Libraries.- G. Phage Antibody Libraries: Wholly Synthetic Monoclonal Antibodies.- References.- 3 Transgenic Approaches to Human Monoclonal Antibodies.- A. Introduction.- B. Competing Technologies for the Generation of Therapeutic Antibodies.- C. Origins of Antibody Diversity.- I. Functional Requirements for a Human Immunoglobulin Transgene.- II. Structure of the Human Immunoglobulin Loci.- 1. The Human ? Light Chain Locus.- 2. The Human ? Light Chain Locus.- 3. The Human Heavy Chain Locus.- D. Transgenic Technology.- I. Pronuclear Microinjection.- II. Embryonic Stem Cells.- III. Transgene Constructs.- 1. Bacteriophage Cloning Vectors.- 2. Plasmid Cloning Vectors.- 3. Yeast Artificial Chromosome Vectors.- E. Immunoglobulin Transgenics.- I. High Level and Cell Type Specific Expression.- 1. Cis-acting Regulatory Sequences.- 2. Transgene Expression.- 3. Human Transgene Constructs.- II. Rearrangement.- 1. Target Sequences.- 2. Immunoglobulin Gene Rearrangements in Transgenic Mice.- 3. Light Chain Junctions.- 4. Heavy Chain Junctions.- 5. Repercussions of Mouse B Cell Environment on Human VDJ Joints.- III. Allelic Exclusion.- 1. Background.- 2. Induction of Allelic Exclusion by Rearranged Transgenes.- 3. Response to Allelic Exclusion by Unrearranged Transgenes.- 4. Alternatives to Direct Feedback Allelic Exclusion.- IV. Primary Repertoire.- V. Intracellular Signaling.- 1. Background.- 2. B Cell Receptor Complex.- 1. Pre-B Cell Complex.- II. Class Switching.- 1. Background.- 2. Class Switching in Transgenic Mice.- 3. Importance of Class Switching for a Human Antibody Mouse.- III. Substrate for Somatic Mutation.- IV. Domination of the Immune Response.- 1. Antibody Depletion.- 2. Anti-sense Transgenes.- 3. Gene Targeting.- A. Perspective.- References.- Section II: Genetically Engineered Monoclonal Antibodies.- 4 Humanization of Monoclonal Antibodies.- A. Introduction.- B. Structure of Antibodies.- I. General.- II. The Antibody Combining Site.- III. Complementarity Determining Regions.- IV. Influence of Framework Residues on Combining Site Structure.- C. Strategies for the Humanization of Antibodies.- I. Transplanting a Nonhuman Combining Site onto a Human Framework.- II. Recombinant Methodology of Complementarity Determining Region Transfer.- 1. Polymerase Chain Reaction-Mediated Complementarity Determining Region Transfer.- 2. Humanization of the Murine Monoclonal Antibody IB4.- 3. Antibody Reshaping.- III. Replacing Surface Residues to Humanize (Veneering).- D. Immunogenicity of Humanized Antibodies.- E. Conclusion.- References.- 5 Applications for Escherichia coli-Derived Humanized Fab' Fragments: Efficient Construction of Bispecific Antibodies.- A. Introduction.- B. Choice of Antigen Specificities for Bispecific F(ab')2.- C. Expression of Humanized Fab' Fragments in E. coli.- D. Recovery of Fab'-SH Fragments.- E. Construction of Bispecific F(ab')2.- F. Uses of E. coli-Derived Fab' Fragments.- G. Conclusions.- References.- Section III: MAb Conjugates and Fusions.- 6 Immunotoxins.- A. Introduction.- B. Considerations in Immunotoxin Development.- I. In Vitro Testing To Identify Effective Antibodies.- II. Immunotoxin Design.- C. The Toxic Moiety.- I. Ricin.- II. Pseudomonas Exotoxin A.- III. Diphtheria Toxin.- IV. Drug Conjugates.- V. Novel Approaches.- D. Cell Biology of Immunotoxin Action.- E. Pharmacology of Immunotoxin Administration.- I. Pharmacokinetics.- II. Pharmacologic Enhancement of Immunotoxin Action.- III. Immunogenicity.- F. Clinical Applications.- I. Cancer.- II. Immunosuppression.- 1. Transplantation.- 2. Autoimmune Disease.- III. Infectious Diseases.- I. Disordered Cellular Growth.- A. Conclusions.- References.- 7 Antibody-Enzyme Fusion Proteins and Bispecific Antibodies.- A. Introduction.- B. Antibody-Enzyme Fusion Proteins.- I. Development of the Concept of a Bifunctional Protein.- II. Chemically Cross-Linked Conjugates as Models for Fusion Proteins.- 1. Cross-Linked Antibody-Plasminogen Activator Conjugates.- 2. Methods for Synthesizing and Purifying Cross-Linked Conjugates.- III. Fusion Protein Construction.- 1. Cloning the Rearranged Immunoglobulin Gene.- 2. Constructing the Expression Vector.- 3. Selecting Loss Variant Cell Lines.- 4. Transfecting the Expression Plasmid.- 5. Purifying and Analyzing Protein.- 6. Recombinant Protein Expression Levels.- IV. Structural and Functional Properties of Specific Fusion Proteins.- 1. Antibody-Plasminogen Activator Fusion Proteins.- 2. A Model Minimal Size Fusion Protein.- 3. Fv and Single Chain Fv.- 4. Minimal Fv-Containing Fusion Protein.- 5. Prodrug Activation.- C. Bispecific Antibodies.- I. Development of the Concept of a Bispecific Antibody.- II. Chemically Cross-Linked Bispecific Antibodies as Models.- III. Cell Fusion in the Production of Bispecific Antibodies.- IV. Functional Properties of Bispecific Antibodies.- D. Conclusion.- References.- 8 Three Generations of Recombinant CD4 Molecules as Anti-HIV Reagents.- A. Introduction.- B. General Aspects of HIV Infection.- C. Characteristics of Different Forms of sCD4.- I. First Generation: Truncated Forms of sCD4.- II. Second Generation: CD4-Immunoglobulins.- III. Third Generation: CD4-FvCD3 Janusins.- D. Molecular Designs and Strategies to Produce Recombinan…