lab 5 - The Immunology Laboratory Introduction to ELISA,...

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The Immunology Laboratory Introduction to ELISA, Hemagglutination and Complement Mediated Lysis When a foreign antigen is introduced into an animal, the animal will respond immunologically to it. The response may be of two different types: cell-mediated or humoral. The effector molecule in the humoral response is immunoglobulin (Ig). The first Ig to appear in a primary immune response is IgM, which is followed in a few days by IgG. The Ig is secreted by antibody forming cells and is specific for the antigen used to immunize the animal. The antibody forming cells are present in the secondary lymphoid organs (spleen, lymph nodes) of the immunized animal. Antibody can be detected in the serum of animals or purified from cells secreting antibody molecules by different in vitro assays. One useful application for antibodies is the e nzyme l inked i mmunos orbent a ssay (ELISA) where specific antibodies are used to label cells within a tissue or cell mixture. Another assay is the agglutination assay (for particulate antigen). If red blood cells are used as a source of antigen the assay is termed hemagglutination. Antibody by itself is rather ineffectual in eliminating foreign organisms. However, antibody of the IgM and IgG class can activate the “complement” (C’) system resulting in a stimulation of different effector functions such as phagocytosis and lysis of foreign organisms. If red blood cells are used as a source of antigen it is relatively simple to demonstrate the lytic properties of the antigen-antibody-complement complex. Complement fixation can also be used as a sensitive method to detect small amounts of antibody. The purpose of this laboratory is to: 1) Demonstrate how the ELISA assay is used to diagnose exposure to a disease by testing for the presence of antibodies to the disease in a sample of simulated serum. An ELISA assay such as the one you will do in this lab would be used to diagnose HIV infection by detecting the presence of
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lab 5 - The Immunology Laboratory Introduction to ELISA,...

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