Figure 6.

Llama-derived anti-candidalysin nanobodies neutralize the hemolytic potential of candidalysin. (a) Candidalysin (4 µM) was pre-incubated with different concentrations of the two nanobody clones (CAL1-F1/CAL1-H1) for 1 h. (b) Melittin (4 µM) was preincubated with 8 µM of the two nanobody clones. The mixture was then added to the purified RBCs, and hemolysis was quantified after 1 h incubation at 37 °C, by measuring the absorbance of sample’s supernatant at 414 nm, and plotted relative to the full lysis control sample (RBCs incubated with pure water), following subtraction of the vehicle control. Each data point on the graph represents a different donor (average of 2 technical replicates). Error bars show the standard deviation. For statistical analysis, an arbitrary value of 0.01 was assigned to any value that was below this threshold. Student’s paired t-tests were then performed on log transformed data. (a) t-test vs. candidalysin only; (b) t-test vs. vehicle control only (^§) or vs. melittin only (^#). *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.001; ns, not significant.