These bound antibodies are then detected using a secondary labeled antibody. In other ELISA formats, the target antigen may be bound directly to the plate and used to detect antibodies present in a sample. This second antibody may be labeled with an enzyme substrate that enables detection, or it may be detected with a labeled species-specific secondary antibody (e.g., anti-human IgG). After thorough washing, a second antibody is added, which binds to the antigen, creating an antibody-antigen-antibody “sandwich”. In a sandwich ELISA, an antibody is bound to the plate surface and used to capture a target antigen from a sample such as plasma or serum. ELISA (Enzyme Linked Immunosorbent Assays) and Western blot techniques are typical examples of commonly used immunoassays.ĮLISAs are performed in multiwell plates in several formats. cdc.gov/hiv/pdf/guidelines_testing_recommendedlabtestingalgorithm.Immunoassays are convenient and widely used detection methods based on the ability of antibodies to bind and detect specific antigens.
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