Method Description

The anti-factor H enzyme-linked immunoassay assay for the quantitation of antibodies to complement factor H is a 3-step procedure. In the first step, standards, controls, and diluted patient specimens are incubated with human recombinant complement factor H immobilized on a microwell plate. During this incubation, antibodies to factor H (AFH) present in the standards, controls, and patient sample will bind to the factor H-coated microwell plate. After incubation, a wash cycle removes the unbound material. In the second step, anti-human IgG conjugated to horseradish peroxidase (HRP) is added to the wells and incubated. The conjugate reacts with the AFH bound to the microwell plate. After incubation, a wash cycle removes the excess conjugate. In the third step, a chromogenic enzyme substrate is added to the wells and incubated. The bound HRP-conjugate reacts with the substrate forming a blue color. The enzyme reaction is stopped by dispensing an acidic solution into the wells, changing the color of the solution from blue to yellow. The color intensity of the reaction mixture is measured spectrophotometrically at 450 nm and is directly proportional to the amount of AFH present in the patient specimens, standards, and controls.(Package insert: Anti-Faktor H. GA Generic Assays GmbH; 05/2022)