B2-OKT3 and huNKG2D-OKT3 binding to rMICA was measured by biolayer interferometry. Streptavidin biosensors were loaded with biotinylated rMICA, saturated with the blocking BiTE (A) huNKG2D-OKT3 or (B) B2-OKT3 and then dipped into the test analytes: anti-MICA/B, huNKG2D-OKT3, B2-OKT3, and buffer only. Data are representative of at least two independent experiments. Cultured human T cells from four different donors were plated with either K562-Luc or Panc1-Luc at a 4:1 ratio. (C) huNKG2D-OKT3 or (D) B2-OKT3 was added at 50ng/ml. rMICA was at the indicated concentrations. ** Indicates concentrations where the cytotoxicity was significantly lower than no rMICA controls, p < 0.01.