Figure 2.
Haemolysis assays. (a) Haemolysis assay on run‐through fractions (each 4 ml) collected from pooled normal human serum (NHS) passed over the RO7112689 column; less than 10% lysis seen on each fraction; complete depletion of human C5. NHS (1 : 10) was used as the positive control (100% lysis). (b) Repletion with C5; C5 sourced either from RO7112689 purification or a commercial source was added back to neat C5‐depleted serum (C5D) at 75 μg/ml; both C5 sources fully restored haemolytic activity but commercial C5 restored serum was less lytic; calculated CH50 values were: RO7112689 C5, 63·4 Units; commercial C5, 40·2 Units. (c) SKY59‐purified C5 or commercial C5 were titrated back into 10% C5D to measure the dose that restored haemolysis to 50%. The 50% lytic dose of RO7112689‐purified C5 was 221·5 ng/ml, whereas that for commercial C5 was 714·7 ng/ml. (d) Titration to identify the lowest lytic dose of SKY59 purified C5 when added back to C5D; C5 at 2 ng/ml conferred measurable lytic capacity in 10% C5D. RO; RO7112689.