Activation of the classical and alternative pathways of complement fixation by immune complexes containing normal and...

Koshland's reagent (2-hydroxy-5-nitrobenzyl bromide (NBB)) has been shown to modify tryptophanyl residues in anti-ovalbumin IgG. As little as 2 moles NBB/mole IgG antibody are sufficient to block the classical pathway of complement activation when the antibody is complexed to antigen (ovalbumin). In contrast, immune complexes containing antibody with the same degree of tryptophanyl substitution will activate the alternative pathway of complement fixation. Immune complexes containing F(ab')2 fragments derived from anti-ovalbumin IgG do not activate the classical pathway. When measuring the percentage activation of C3 using the method of Laurell, NBB does not affect the alternative pathway of the complement system up to a molar ratio of 2 NBB/F(ab')2. The above findings, provide a means to evaluate the relative contribution of complement activation by the different pathways.