Fig. 4: Hexamerization-enhancing mutation increases complement activation capacity of anti-capsule antibodies. | Nature Communications

Fig. 4: Hexamerization-enhancing mutation increases complement activation capacity of anti-capsule antibodies.

From: Agnostic B cell selection approach identifies antibodies against K. pneumoniae that synergistically drive complement activation

Fig. 4

KpnO2 was incubated with (a) anti-O2 antibodies (UKpn2 and UKpn6) as wildtype and E430G variant or (b) anti-capsule antibodies (UKpn1 and UKpn3) as wildtype and E430G variant or for 30 min at 4 °C. After antibody binding, bacteria were incubated with 3% KpnO2∆NHS as a complement source for 30 min at 37 °C. C3b deposition was detected using anti-hu-C3b-A647 and analyzed by flow cytometry. c KpnO1 was incubated with the antibodies UKpn72, UKpn68 or UKpn82 as wildtype and E430G variant for 30 min at 4 °C. After antibody binding, bacteria were incubated with 1% KpnO1∆NHS as a complement source for 30 min at 37 °C. C3b deposition was detected using anti-hu-C3b-A647 and analyzed by flow cytometry. a–c Flow cytometry data are represented by gMFI of bacterial populations. Data represent mean ± SD of three independent experiments.

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