The IAA-ELISA is an enzyme-linked immunosorbant assay (ELISA) for IAA determination.
Human insulin is immobilized onto microwells. The reference, positive, and negative controls, and diluted patient serum samples are added to the appropriate microwells. Human IgG specific antibodies to insulin in the serum sample and controls bind to the insulin molecules on the microwells. After washing off unreacted serum materials,
an enzyme (alkaline phosphatase) labelled goat antibody specific to human IgG is added to the antigen-antibody complex. After thorough washing to remove the unbound enzyme, a substrate (PNPP) solution is added and the color development is measured spectrophotometrically. The intensity of the color is directly proportional to the concentration of IAA in the sample. Two quality controls (positive and negative) are provided to monitor and validate assay results.