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Monoclonal Antibodies Methods and Protocols 2nd Edition by Vincent Ossipow, Nicolas Fischer ISBN 1493963228 9781493963225

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ISBN 10: 1493963228
ISBN 13: 9781493963225
Author: Vincent Ossipow, Nicolas Fischer 

Monoclonal Antibodies: Methods and Protocols, Second Edition expands upon the previous edition with current, detailed modern approaches to isolate and characterize monoclonal antibodies against carefully selected epitopes. This edition includes new chapters covering the key steps to generate high quality monoclonals via different methods, from antigen generation to epitope mapping and quality control of the purified IgG. Chapters are divided into four parts corresponding to four distinct objectives. Part I covers monoclonal antibody generation, Part II deals with monoclonal antibody expression and purification, Part III presents methods for monoclonal antibody characterization and modification, and Part IV describes selected applications of monoclonal antibodies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Monoclonal Antibodies: Methods and Protocols, Second Edition provides crucial initial steps of monoclonal antibody generation and characterization with state-of-the art protocols.

Monoclonal Antibodies Methods and Protocols 2nd Table of contents:

Part I: Introduction to Monoclonal Antibodies

  1. History and Discovery of Monoclonal Antibodies

    • Early Development and Milestones
    • Impact on Biomedicine and Biotechnology

  2. Principles and Technology Behind Monoclonal Antibodies

    • The Hybridoma Technique
    • Antibody Structure and Function
    • Sources of Monoclonal Antibodies

  3. Monoclonal Antibodies in Research and Medicine

    • Diagnostic Applications
    • Therapeutic Applications: From Development to Approval
    • Ethical, Legal, and Regulatory Considerations

Part II: Generation of Monoclonal Antibodies
4. Immunization Protocols for Antibody Production

  • Selection of Antigens
  • Immunization Strategies and Adjuvants
  • Optimizing Immune Response

  1. Hybridoma Technology and Screening

    • Fusion of B Cells with Myeloma Cells
    • Selection and Cloning of Hybridomas
    • Screening for Specificity and Affinity

  2. Alternative Technologies for Antibody Generation

    • Phage Display Libraries
    • Single-Cell Technology
    • Transgenic Mice and Alternative Platforms

Part III: Characterization of Monoclonal Antibodies
7. Screening for Antibody Specificity and Affinity

  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Western Blotting and Immunohistochemistry
  • Flow Cytometry for Cellular Analysis

  1. Biophysical and Biochemical Characterization

    • Determining Antibody Binding Kinetics
    • Surface Plasmon Resonance (SPR)
    • Protein-A Chromatography and Mass Spectrometry

  2. Assessment of Functional Properties

    • Antibody-Dependent Cellular Cytotoxicity (ADCC)
    • Complement-Dependent Cytotoxicity (CDC)
    • Neutralization Assays

Part IV: Production and Purification of Monoclonal Antibodies
10. Cell Culture Techniques for Large-Scale Production

  • Optimizing Hybridoma Culture
  • Suspension Cultures and Bioreactors
  • Automation and High-Throughput Screening
  1. Purification and Quality Control
  • Chromatographic Methods (Protein A, Ion-Exchange, etc.)
  • Purification of Antibody Fragments
  • Quality Control and Regulatory Guidelines
  1. Scaling Up: From Laboratory to Commercial Production
  • Process Optimization and Yield Maximization
  • Good Manufacturing Practices (GMP)
  • Regulatory and Compliance Requirements

Part V: Engineering Monoclonal Antibodies
13. Antibody Humanization and Chimerization

  • Methods of Humanizing Monoclonal Antibodies
  • Reducing Immunogenicity and Improving Efficacy
  1. Designing Antibody Fragments and Bispecific Antibodies
  • Fab, scFv, and Nanobody Formats
  • Engineering Bispecific and Trispecific Antibodies
  • Applications of Antibody Fragments
  1. Antibody Engineering for Targeted Therapy
  • Conjugating Monoclonal Antibodies to Drugs or Toxins
  • Antibody-Drug Conjugates (ADCs)
  • Antibodies in Gene Editing: CRISPR-Cas9 Applications

Part VI: Applications of Monoclonal Antibodies
16. Monoclonal Antibodies in Cancer Therapy

  • Targeted Therapy and Immuno-Oncology
  • Immune Checkpoint Inhibitors
  • CAR-T Cells and Monoclonal Antibodies
  1. Monoclonal Antibodies in Infectious Disease
  • Antibodies Against Viruses (HIV, SARS-CoV-2, etc.)
  • Monoclonal Antibodies for Bacterial Infections
  • Vaccine Development and Antibody Protection
  1. Monoclonal Antibodies in Autoimmune and Inflammatory Diseases
  • Mechanisms of Action in Autoimmunity
  • Antibodies in Rheumatoid Arthritis, Crohn’s Disease, and More
  1. Diagnostic and Imaging Applications
  • Monoclonal Antibodies in Diagnostic Testing (ELISA, Lateral Flow, etc.)
  • Antibody-Based Imaging in Medical Imaging and Diagnostics

Part VII: Emerging Trends and Future Directions
20. Advances in Antibody Engineering and Therapeutics

  • Next-Generation Monoclonal Antibodies
  • Bispecific T-cell Engagers (BiTEs)
  • Immunoediting and Antibody Optimization
  1. Novel Platforms and Alternatives to Monoclonal Antibodies
  • Nanobodies and VHHs
  • Engineered Antibody Alternatives: Aptamers and Scaffolds
  • Advances in Antibody-Drug Conjugates (ADCs)
  1. Challenges and Opportunities in Monoclonal Antibody Development
  • Overcoming Resistance and Side Effects
  • Overcoming Manufacturing Challenges
  • The Future of Antibody-Based Therapies in Precision Medicine

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