Fig. 2.

The purified anti-CFH autoantibody. a Confirmation of purified anti-CFH autoantibodies from our patient by Western blot. Commercial human CFH (1 μg) under non-reducing conditions was electrophoresed on 10% SDS-PAGE and transferred to a PVDF paper, blocked and then incubated with anti-CFH autoantibodies purified from our patient. The anti-CFH autoantibodies purified from our patient, which was IgG3λ, could recognize commercial CFH. Lanes a-e were incubated with the followings antibodies: anti-human IgG, IgG1, IgG2, IgG3, and IgG4, respectively. b The anti-CFH autoantibodies purified from our patient had lambda light chain without a kappa chain. Lanes f and h, lanes g and i were loaded with anti-CFH autoantibodies purified from the aHUS patient and our patient, respectively, under non-reducing conditions on 8% SDS-PAGE; lanes f and g were then incubated with anti-human kappa chain, and lanes h and i were incubated with anti-human lambda chain. c The serum of our patient contained a monoclonal IgGλ by immunofixation electrophoresis assay. d The purified anti-CFH autoantibodies from our patient were confirmed to be the same monoclonal IgGλ by immunofixation electrophoresis assay