C1q binding to surface-bound IgG is stabilized by C1r2s2 proteases

Zwarthoff, Seline A.; Widmer, Kevin; Kuipers, Annemarie; Strasser, Jürgen; Ruyken, Maartje; Aerts, Piet C.; de Haas, Carla J.C.; Ugurlar, Deniz; Vidarsson, Gestur; Van Strijp, Jos A. G.; Gros, Piet; Parren, Paul W.H.I.; van Kessel, Kok P.M.; Preiner, Johannes; Beurskens, Frank J.; Schuurman, Janine; Ricklin, Daniel; Rooijakkers, Suzan H.M.

Complement is an important effector mechanism for antibody-mediated clearance of infections and tumor cells. Upon binding to target cells, the antibody’s constant (Fc) domain recruits complement component C1 to initiate a proteolytic cascade that generates lytic pores and stimulates phagocytosis. The C1 complex (C1qr2s2) consists of the large recognition protein C1q and a heterotetramer of proteases C1r and C1s (C1r2s2). While interactions between C1 and IgG-Fc’s are believed to be mediated by the globular heads of C1q, we here find that C1r2s2 proteases affect the capacity of C1q to form an avid complex with surface-bound IgG molecules (on various DNP-coated surfaces and pathogenic Staphylococcus aureus). The extent to which C1r2s2 contribute to C1q-IgG stability strongly differs between human IgG subclasses. Using antibody engineering of monoclonal IgG we reveal that hexamer-enhancing mutations improve C1q-IgG stability, both in absence and presence of C1r2s2. In addition, hexamer-enhanced IgGs targeting S. aureus mediate improved complement-dependent phagocytosis by human neutrophils. Altogether, these molecular insights into complement binding to surface-bound IgGs could be important for optimal design of antibody therapies.