Figure 1.

Strategy used to select antibodies using both phage and yeast display. Antibodies are first selected against mycolactone using two rounds of phage display, after which the whole selection output is cloned into a yeast display vector. A further one or two rounds of sorting by flow cytometry allow the subsequent isolation and testing of single clones, followed by affinity maturation by mutagenesis to select for higher affinity binders. The final selected antibodies are finally expressed as ‘scFv-Fc fusions’—where the variable domain of the antibody (scFv) is fused with the CH2-CH3 constant region (Fc) of human immunoglobulin IgG1—and further validated in an enzymatic assay (ELISA) for its specific binding to the toxin of interest.