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. 2019 Jun 17;157(4):283–295. doi: 10.1111/imm.13083

Figure 2.

Figure 2

Direct and sandwich ELISA for characterization of new monoclonal antibodies (mAb). In direct ELISA, plates were coated with human C5 (a), C5b6 (b) or C6 (c). All new mAb detected C5 and C5b6 complex, but not C6. In a sandwich ELISA with RO7112689 as the capture and in house 4G2 mAb as detection (d), both C5 and C5b6, but not C6 were detected showing that mAb 4G2 and RO7112689 antibodies recognized different epitopes in C5. Sandwich ELISA comparing RO7112689 (e; anti‐C5β) and Eculizumab (f; anti‐C5α) as capture mAb showed that both the C5α‐specific mAb (7D4 and 4G2) bound strongly to RO7112689‐captured C5 but the C5β‐specific 10B6 did not bind (e); the β‐chain‐specific mAb 10B6 bound most strongly to Eculizumab‐captured C5 whereas the anti‐C5α mAb bound weakly (4G2) or not at all (7D4), suggesting that they competed for similar epitopes (f). All experiments were repeated three times with the same results. The error bars are standard errors of triplicates.