Assay Information
Highly purified antigens SSA (Ro), SSB (La), Sm RNP/Sm, Scl70 and Jo1 are bound to microwells individually. Antibodies against these antigens, if present in diluted serum or plasma, bind to the respective antigen. Washing of the microwells removes unspecific serum and plasma components. Horseradish peroxidase (HRP) conjugated antihuman IgG immunologically detects the bound patient antibodies forming a conjugate/antibody/antigen complex. Washing of the microwells removes unbound conjugate. An enzyme substrate in the presence of bound conjugate hydrolyzes to form a blue color. The addition of an acid stops the reaction forming a yellow endproduct. The intensity of this yellow color is measured photometrically at 450nm. The amount of colour is directly proportional to the concentration of IgG antibodies present in the original sample.