Complement deposition was performed as described before (45 (link)). In brief, biotinylated antigens were coupled to fluorescent neutravidin beads (Thermo Fisher) and incubated with 10 μl of 1:10 diluted plasma. Beads were incubated with guinea pig complement in GVB++ buffer (Boston BioProducts, MA, USA) for 20 min at 37°C. Deposited C3 on beads was stained with anti-guinea pig C3 antibody labeled with fluorescein isothiocyanate (FITC) (MP Biomedicals, CA, USA) and analyzed by flow cytometry on a LSRII instrument (BD Biosciences, CA, USA).
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