Figure 2.
Single-domain antibody characterization. (A) Flow cytometry analysis of the different phage clones using SKOV3 cells. A total of 5×105 SKOV3 cells were used for per sample, and 1 μg of purified phage binders and 1 μg M13 Ms mAb were used as the primary antibodies. Goat anti-mouse IgG-AF488 used as the detection antibody. For simplicity, only clones B3, E6, and C3 are shown. NC refers to negative control, and ISO refers to isotype. (B) The relatively strong binders were selected for the ELISA with a serial dilution of the Her2 antigen. (C). Ranking of the VHHs based on the flow cytometry analysis, as described in Figure 2A; 10 of the clones (gray bars) showed an obvious shift, and 5 of them were further analyzed (gray bars with black dots).