FIGURE 1.

Camelid immunization combined with YSD allows for the generation of a diverse panel of NKp46‐specific sdAbs for NK cell redirection. (a) Schematic depiction of a bispecific SEEDbody for NK redirection based on a NKp46‐specific VHH molecule in combination with a humanized version of the Fab arm of Cetuximab for EGFR targeting. The scheme was generated using PyMol software version 2.3.0. (b) FACS‐based selection for the isolation of NKp46‐specific VHHs by utilization of a two‐dimensional staining strategy for full‐length VHH display and NKp46 binding at a concentration of 1 μM. Of note, plots in the upper row show representative 10⁶ cells of the sort, while plots in the lower row show representative 5 × 10⁴ cells of the corresponding sorting output in order to visualize enrichment. (c) Graphical alignment of unique amino acid sequences of 40 VHH clones obtained from YSD library sorting. CDRs are indicated. Red bars represent high sequence diversity and green bars indicate high sequence conservation at a given position. Alignment obtained using MUSCLE alignment with Geneious Prime® January 1, 2021 software.