Extended Data Fig. 1: CDH17 as the antigen of VHH1 nanobody is expressed in NET BON cells.
(a) Flow cytometry analysis of VHH1 nanobody binding to various cell lines. Representative of n = 2 independent experiments with similar results. (b) Western Blot analysis of CDH17 protein expression in various cell lines. ACTIN as a control. Experiments were performed two independent times with similar results. (c) 293 T cells were transfected with full length CDH17 or truncation mutant plasmids, followed by Western Blot analysis of CDH17 protein expression by using VHH1 or commercial anti-CDH17 antibody (Santa Cruz, clone H-1) which binds to the C-terminal domain of CDH17. ACTIN as a control. Experiments were performed two independent times with similar results. (d-e) SPR detection of the binding affinity of the CDH17 protein to the VHH1 nanobody. The equilibrium dissociation constant KD of the CDH17 protein to the VHH1 nanobody was 4.79 × 10-7 M.
