Abstract
Abstract The standard method for hemolytic titration of C2 involves a stepwise build-up of cellular intermediates in the complement sequence. The requirements for purified hemolytically active components and cellular intermediates make this assay difficult to perform. As sera from patients with selective deficiency of C2 contain all the other complement components in normal titers, these sera serve as ideal reagents for a one-step hemolytic titration for C2 activity. Accordingly, we have analyzed the optimal conditions of the variables which affect C2 titration in a one-step assay and have compared its sensitivity to that of the standard method. In initial studies, a reaction mixture consisting of equal volumes (0.2 ml) of EA (5 × 108 cells/ml), C2D (1:20) (Clin. Immunol. Immunopathol. 4:269, 1975) and functionally purified C2 (Cordis Laboratories), produced lysis which was found to be dependent on the incubation time and concentration of C2.
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