Basic methods in antibody production and characterization Routledge Revivals Series

Written for researchers and professionals in the fields of biomedical research, immunology, biochemistry, molecular biology, pathology, and biotechnology, Basic Methods in Antibody Production and Characterization uses a cookbook approach to presenting the methods for the production, characterization, and use of antibodies. Antibodies described include polyclonal and monoclonal and those made by the phage display. Basic Methods in Antibody Production and Characterization focuses on specific aspects of antibodies and takes readers on a step-by-step process from antigen preparation to immunizations, adjuvants, screening, purification, storage, and applications.
Introductory material accompanies each chapter, giving readers a better understanding of the methods and applications. Ample references arm researchers with many applications pertaining to the particular technology.

Starting out, delia r. bethell and gary c. howardMonoclonal vs. Polyclonal Antibodies Using This Manual Laboratory Safety Animal welfare and regulatory issues, sigmund t. richIntroduction Legal Aspects Compliance Problems Compliance Solutions Synopsis of the Animal Welfare Act Antigens, donald h. lewisIntroduction Haptens, Immunospecificity, and Immunogen-Antibody Interactions Specificity and Cross-Reactivity Peptides As Immunogens Potential Pitfalls Adjuvants, jory r. baldridge and michael j. lacyIntroduction General Guidelines and Precautions for Working with Adjuvants Protocols for Using Freund's Adjuvants Protocol for Using Titermax7 Use of Ribi Adjuvant System Polyclonal antibodies, eric s. beanIntroduction Selection of Host Injection Protocols Dose of Antigen Choice of Adjuvant Route of Immunization Bleeding Testing Serum Samples Post-Production Purification Monoclonal antibody production, sandy j. stewartIntroduction Cell Lines and Media Immunization Hybridization Protocol Culture and Propagation Cloning and Cryopreservation Screening techniques, gamal e. osman and warren e. ladigesIntroduction Antibody-captured elisa to detect specific antibodiesDouble antibody-sandwich elisaAntibody Capture on Cell Surface Antibody production by ascites growth, kristi r. harkinsIntroduction Ascites Production in Mice (Monoclonal Antibody) Ascites Production in Mice (Polyclonal Antibody) Ascites Production in Rats (Monoclonal Antibody) Growth of cell lines in bioreactors, beverly j. norris, michael j. gramer, and mark hirschelIntroduction Common Bioreactor Themes Batch Culture Fed Batch Culture Perfusion Culture Hollow-Fiber Bioreactor Summary: Choosing a Technology Phage display, peter amersdorfer and james d. marksIntroduction Generation of Human and Murine scFv Gene Repertoires Library Construction Panning Identification of Antigen Binding Clones Expression and Purification of Native Antibody Fragments Antibody purification methods, kristi r. harkinsIntroduction Preliminary Steps Purification IgG Purification of IgM Purification of IgY (Chicken Yolk) Other Methods Final Product Molecular characterization of immunoglobulin genes, warren ladiges and gamal e. osmanIntroduction Cloning immunoglobulin V Regions Using PCR with Universal Primers Cloning Ig V Regions Using Linkered-Ligated PCR Antibody storage, kathryn elwell and delia r. bethellIntroduction Storage of Serum, Ascites, and Tissue-Culture Supernatant Storage of Purified Antibodies Storage of Conjugated Antibodies Chemically modifying antibodies, kathryn elwell and gary c. howardBasic Considerations Sites for Modification Adding a Fluorescent Label Adding Biotin to Amino Groups Activating carboxyl groups with edacAdding Biotin to the Carbohydrate Group Radioiodination by the Bolton-Hunter Procedure Radioiodination by the Chloramine T Procedure Bifunctional Reagents and Conjugations Cross-Linking with Glutaraldehyde Preparing antibody-enzyme conjugates with spdpPreparing Antibody-Enzyme Conjugates with MBS Preparing antibody-enzyme conjugates with smccPreparing Horseradish Peroxidase-Antibody Conjugates Preparing Antibody-Phycobiliprotein Conjugates Western blotting, lee bendickson and marit nilsen-hamiltonIntroduction Transferring Proteins Antigen Detection Artifacts and Troubleshooting Clinical Applications Conclusion Western Blot Protocol Immunohistochemical methods, kendrick j. morrell, neil m. hand, and nicholas r. griffinIntroduction Tissue Preparation Enzyme Labels Achieving Specificity Pretreatment Techniques Immunoenzyme Techniques The Indirect Peroxidase Technique The Labeled Streptavidin/Biotin (LSAB) and the Streptavidin Biotin Complex (ABC) Techniques The Alkaline Phosphatase Anti-Alkaline Phosphatase (APAAP) Technique Immunoflourescence Techniques The Direct Immunoflourescence Technique Specialized Techniques

Researchers, students in biochemistry, molecular biology, immunology, biology, zoology, and related fields