The objective of this lecture and laboratory course is to provide investigators with information on chemical approaches to the isolation, purification and characterization of antibodies and antigens. Special emphasis will be given to monoclonal antibody formation, assay, and characterization.
The Immune Response; Immunoglobulin Structure and Diversity; Immunogenetics; Antigenic Determinants; Idiotypic Network; Application of Monoclonal Antibodies; Cell Fusion; Myeloma Cell Lines; Screening for Monoclonal Antibodies (ELISA, Immunofluorescence, RIA); Cloning; Purification and Characterization Approaches; Human Monoclonal Antibody Formation and use; Crystal Structure; Purification of Antibodies from Serum, Ascites, and Culture Supernatant; Analysis of Antibody-Antigen Reactions by Immunoprecipitation and Equilibrium Dialysis; Affinity Determinations; Production of Idiotypic Antibodies, Antibody Fragments, and Novel Antibodies; Peptide Carriers and Antigen Synthesis; Radioreceptor and Crosslinking Assays for Cell Surface Receptors; Immunocytochemistry; Recombinant DNA Approaches.
Course emphasis is placed on understanding the immune system principles and the core immunology research methods. Particular focus will be placed on the cellular elements and their roles in the orchestration of the immune response. Because this field is contributing to novel therapies and is in a high state of flux, due attention will be given to new directions.
Cells of the adaptive immune system and of the innate immune system; special focus on B lymphocytes and T lymphocytes; Interactions between antibodies and antigens; Genes encoding antibodies; Immunological memory and the mechanisms involved in recall responses; The Major Histocompatibility complex (MHC) and how it controls adaptive immune response; Immunological tolerance and how the immune system is kept under control; Immunotherapy; How to generate and produce monoclonal antibodies; Using a flow cytometer for the analysis of cells of the immune system; Immunological assays (T cell proliferation, cytotoxic T lymphocyte cell assays, Western Blotting/Immunostaining, ELISA); The Mucosal Immune System.
Immunocytochemistry is an important research tool which has significant applications in topics as important and diverse as basic cell biology, carcinogenesis, apoptosis, genomics, clinical diagnosis, and intracellular migration and localization of molecules. This 3 day lecture and lab course will introduce the participants to the key methods and applications of immunocytochemistry and their underlying principles.
Introduction to Immunofluorescence; Conjugation of Proteins with Fluorochromes; Tissue Preparation for Immunocytochemistry; Immunocytochemistry on Whole Mount Preparations; In situ Hybridization with Immunoprobes; Labeling and Flow Analysis of Cells; Applications of Flow Cytometry and Confocal Microscopy. Preparation of and Fluorescent Labeling of Cultured Cells; ABC-Peroxidase and Acid Phophatase Labeling of Sectioned Material; ELISA on Fixed Cells; Fluorescein Labeling of Nerve Cells in Whole Mount.