EVALUATION OF COMPLEMENT ACTIVITY BY AN ENZYME-IMMUNOASSAY

An ELISA-type assay useful for the evaluation of the complement activity in serum is described. Aggregated pooled human IgG (IgGn) prepared so as to exclude large and small aggregates, to resemble soluble circulating immune complexes, was used to coat polystyrene microwells to serve as initiator of complement activation. Fresh serum, at different dilutions, as the source of the complement to be evaluated was added and the plate incubated 90 min at 37-degrees-C. Then, a peroxidase-labeled antihuman C3c antibody was added to react with the bound C fragments. This was followed by 2,2'-azino-di-3-ethyl benzothiazoline sulfonic acid (ABTS), as the color reagent used for detection of the enzyme activity. In this system, theoretically, the levels of activating and regulatory complement components are evaluated up to the level of C3 splitting. The assay was applied in healthy volunteers to set normal values and in 15 patients suffering from systemic lupus erythematosus making possible the differentiation of those with normal and low complement levels.