Monoclonal Antibody Protocols

1995
ISBN: 978-0-89603-308-5
Verlag: Humana Press

Buch, Englisch, Band 45, 264 Seiten, Format (B × H): 155 mm x 235 mm, Gewicht: 474 g

Reihe: Methods in Molecular Biology

Since the initial description of techniques to immortalize anti­ body-producing B-lymphocytes by fusion with tissue culture-adapted myeloma cells, methods have been developed to produce monoclonal antibodies of defined specificity in multiple animal species. Stable hybrids can be readily produced in mice using a number of myeloma and hybridoma cell lines. To obviate the problem of identifying fusion partners in other animal species, xenohybrids have been produced using B-lymphocytes from the relevant species and mouse myeloma cells. The use of xenohybrids has minimized the problem of obtain­ ing stable antibody-producing hybrids in all species examined thus far. Although alternative techniques are being developed to produce monoclonal antibodies by molecular methods, hybridoma technol­ ogy will remain the technology of choice for producing monoclonal antibodies for a variety of applications in research and industry. The objective of Monoclonal Antibody Protocols is to provide investigators with a set of methods for producing and using mono­ clonal antibodies in biomedical, agricultural, and biological sciences. The book is not intended to provide methodology for all possible applications, but rather a series of methods presented in an easy-- follow format that can be used by new and established investiga­ tors, graduate and postgraduate fellows, and technical staff.

Zielgruppe

Research

Autoren/Hrsg.

Davis, William C.

Fachgebiete

• Naturwissenschaften Biowissenschaften Tierkunde / Zoologie Wirbeltiere (Vertebrata) Säugetiere (Mammalia) Meeres- und Süßwassersäuger
• Medizin | Veterinärmedizin Medizin | Public Health | Pharmazie | Zahnmedizin Klinische und Innere Medizin Immunologie
• Naturwissenschaften Biowissenschaften Tierkunde / Zoologie Wirbeltiere (Vertebrata) Säugetiere (Mammalia) Beuteltiere (Monotremen)
• Naturwissenschaften Biowissenschaften Tierkunde / Zoologie Wirbeltiere (Vertebrata) Säugetiere (Mammalia) Primaten
• Naturwissenschaften Biowissenschaften Proteinforschung
• Naturwissenschaften Biowissenschaften Molekularbiologie

Inhaltsverzeichnis

Methods of Immunization to Enhance the Immune Response to Specific Antigens In Vivo in Preparation for Fusions Yielding Monoclonal Antibodies.- Methods of Immunization to Enhance the Immune Response to Specific Antigens In Vitro.- Culture Conditions that Optimize Outgrowth of Hybridomas.- Production of Stable Heterohybridomas Producing Human Monoclonal Antibodies.- Production of Stable Bovine-Murine Interspecies Hybrids.- Production of Monoclonal Antibodies in Swine.- Production of Monoclonal Antibodies in Horses.- Use of Electric-Field-Mediated Cell Fusion to Produce Hybridomas Secreting Monoclonal Antibodies.- Rapid Isotyping of Mouse Monoclonal Antibodies.- Methods of Measuring Antibodies in Sera/Plasma, Ascites, and Tissue-Culture Media.- Measurement of Immunoglobulin Synthesis Using the ELISPOT Assay.- Use of Monoclonal Antibodies in Immunohistochemistry.- Use of Monoclonal Antibodies in Immunoelectron Microscopy.- Use of Monoclonal Antibodies for Western Blotting with Enhanced Chemiluminescent Detection.- The Use of Directly and Indirectly Labeled Monoclonal Antibodies in Flow Cytometry.- Use of Monoclonal Antibodies and Flow Cytometry to Cluster and Analyze Leukocyte Differentiation Molecules.- In Vivo and In Vitro Production of Monoclonal Antibodies.- Separation of Monoclonal Antibodies from Cell-Culture Supernatants and Ascites Fluid Using Thiophilic Agarose.- Detection, Purification, and Utilization of Murine Monoclonal IgM Antibodies.- Preparation and Use of Immunoaffinity Columns with Monoclonal Antibodies Without Purification from Ascites and Tissue-Culture Medium.- Antigen Purification by Monoclonal Antibody Immunoaffinity Chromatography.- Coupling of Monoclonal Antibodies with Fluorophores.- Coupling of Monoclonal Antibodies with Biotin.- Coupling of Monoclonal Antibodies with Enzymes.- Use of Monoclonal Antibodies with Magnetic Particles to Separate Cell Subpopulations by Negative Selection.- Use of Monoclonal Antibodies with Magnetic Particles to Separate Cell Subpopulations by Positive Selection.