Figure 4. Mapping of the epitope specificity of nanobodies by cross-blocking analyses.

For analysis of epitope specificities, selected nanobodies were subjected to competitive binding analyses (A). Antigens were immobilized on 96-well plates and pre-incubated with nanobody-Fc fusion proteins (2 µg/well) for 60 min at RT before addition of c-myc tagged nanobodies precomplexed with a peroxidase-conjugated anti-c-myc mAb (50 ng nanobody/well). The schematic diagram illustrates the experimental set-up, PO: peroxidase. (B) Schematic diagrams illustrating the relative localization of the binding epitopes of the analyzed nanobodies.