Biotechnology and Genetic Engineering
Monoclonal Antibodies Application in Disease and Health
Description
India is likely to become a major force in the area of Biotechnology in the next decade. The Department of Biotechnology has encouraged human resource development by creating new institutions and centres of excellence for the research and training. Any research work in biotechnology uses Hybridoma Technology and Antibody Engineering as a basic tool in manipulation of cell organelle genetically as well as phenotypically. This book is an attempt to comprehensively present the information on Hybridoma Technology and Antibody Engineering in single volume. It is intended primarily as a text book for undergraduate and post graduate students majoring in Biotechnology, Biosciences, Biotechnology product formulations, preparation of vaccines and diagnostics used in veterinary and human medicine. The new biology is being inducted through animal cell culture and hybridoma management into animal industry. This book has eleven chapters that deal with historic developments, laboratory design, sterilization procedures and various facets of hybridoma culture. This includes preservation, characterizations, storage and transport of cells, their monitoring and technologies for species cell culture.
Table of Contents
About the Series; Preface; Introduction: History of hybridoma technique; Principles of monoclonal antibody production of predefined specificity; Monoclonal antibodies–an overview; On proteins vs nucleic acids; Structure of proteins; DNA and protein electrophoresis; Antigens and antibodies–structure and function-a recap; Antigen–antibody interactions; Requirements of a Hybridoma Laboratory: Introduction; What is cell culture?; Mice facility; Production of Monoclonal Antibodies: Introduction; Collection of spleen; Myeloma cell preparation ; Fusing agents; Components of HAT medium; Fusion of spleen lymphocytes and myeloma cells; Cloning by limiting dilution; Hybridoma cells; Taggert hybridoma technology; Naming the hybridoma clones; Immunoassays for screening of mAbs; Neutralization assays; Radio immuno assay; Characterization of Monoclonal Antibodies: Polyacrylamide gel electrophoresis; Isoelectric focusing; Autoradiography and fluorography; Western blotting; Specificity of the monoclonal antibodies; Isotyping of monoclonal antibodies; Affinity of the mAbs; Characterization of the mAbs for intended use; Selection of mAbs; Mass Production of Monoclonal Antibodies: Expansion of the hybridomas secreting monoclonal antibodies; In vivo production of mAbs; In vitro production of mAbs; Serum-free media; Purification of Monoclonal Antibodies: Introduction; Total protein estimation methods; Proteolysis and its prevention; Purification strategy; Concentration by precipitation; Chromatography; Biomimetic ligands; Covalent affinity chromatography; Magnetic affinity beads; Protein tags; Hydrophobic interaction chromatography; Thiophilic interaction chromatography; Purification of mAbs; Purification of mAbs used for various application; Purity testing; Protein storage; Cryopreservation of Hybridomas: Introduction; Cryopreservation; Principle of cryopreservation; Liquid nitrogen; Programmable freezers; Thawing; Selection of liquid nitrogen containers; Monitoring and replenishing liquid nitrogen; Transportation of cells; Protocol for freezing and thawing of hybridomas; Conjugation of Monoclonal Antibodies: Introduction; Antibody modification sites; Radiolabeling of mAbs; Conjugation of mAbs to fluorescent dyes; Cross linking agents; Enzyme coupling of mAbs; Coupling of mAbs with biotin; Coupling to magnetic particles; Detection methods for coupled mAbs; Anti-tumour therapy using mAbs; Application of Monoclonal Antibodies: Introduction; Antigen antibody interactions–a recap; Application of mAbs in diagnosis; mAbs in immunoassays; Performance characteristics of immunoassays; Application of mAbs in antigenic characterization of viruses; Application of mAbs in detection of antibodies; Application of mAbs in detection of antigen (virus protein); Application of mAbs in detection of antigen (virus protein based quality control of vaccine; Application of mAbs in detection of antibody/antigen to viruses having multiple serotypes; Application of mAbs in differentiation of vaccinated from infected animals (DIVA); Application of mAbs in vivo depletion of T cells; Application of mAbs in aflatoxin detection; mAbs to identify receptors for molecules; mAbs in interferon quantification and diagnosis of Mycobacterial diseases; mAbs in immunophenotyping; Application of mAbs in measuring immunoglobulin levels; mAbs and stem cells; mAbs in pregnancy diagnosis; Anti-idiotype mAbs as vaccines; mAbs in bacterial and parasitic diseases; mAbs in immunochemistry; mAbs in immunoaffinity purification; Application of mAbs as therapeutics; Engineering Fc region of the mAbs; Antibody drug conjugates; Radioimmunotherapy for infectious diseases using mAbs; Single domain antibodies; Affibody; Ribosome display technology; mAbs in imaging; Intrabodies; Fc fusion proteins; Alternate hosts for mAb production; Antibody Engineering: Introduction; Antibody genetics; Humanized antibodies; Prokaryotic systems; Eukaryotic systems; Recombinant expression of antibody fragments; Therapeutic antibodies; Alternatives to Monoclonal Antibodies: Introduction; Production of recombinant antibodies by phage display technology; Manipulating structural proteins of filamentous phages for antibody libraries; Production of mono-specific sera using recombinantly expressed proteins; Steps involved in the overexpression of recombinant proteins; Production of mono-specific serum against recombinant proteins; Anti-peptide antibodies; Plantibodies; References; Suggested reading; Glossary; Appendix; Index