Method Description

This enzyme-linked immunosorbent assay (ELISA) method detects and measures serum levels of antibodies of certain pemphigoid diseases. Calibrators and patient sera are added to microwells coated with collagen type VII antigens, allowing antibodies to react with the immobilized antigens. After washing to remove any unbound serum proteins, horseradish peroxidase-conjugate IgG is added and incubated. Following another wash step, the peroxidase substrate is added and allowed to incubate for an additional period. A stop solution is then added to each well to cancel the enzyme reaction and to stabilize the color development. The assay can be quantified by measuring the reaction photometrically and plotting the results. The amount of antigen-specific bound antibody is proportional to the color intensity.(Unpublished Mayo method)