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. 2021 Dec 13;28(5):948–959. doi: 10.1158/1078-0432.CCR-21-1798

Figure 5.

Figure 5. Humanized AR9.6 demonstrates robust in vitro, in vivo, and ex vivo binding to MUC16-expressing ovarian and pancreatic cancer cells. A, Humanization of muAR9.6 by CDR grafting. B, In vitro internalization data of [89Zr]Zr-DFO-huAR9.6 in OVCAR3 cells at 1, 3, 6, and 24 hours. The blue bars represent the uptake of 10 ng of [89Zr]Zr-DFO-huAR9.6 at each timepoint, whereas the red bars represent the blocked uptake of [89Zr]Zr-DFO-huAR9.6 in the presence of a 1,000-fold excess of unlabeled huAR9.6. C, In vitro and ex vivo validation of the binding of huAR9.6 to MUC16high (OVCAR3) and MUC16neg (SKOV3) ovarian cancer cells using flow cytometry and immunohistochemical (IHC) staining of FFPE tumor sections. D, Ex vivo validation of the binding of huAR9.6 to FFPE sections from MUC16high (S2-028), MUC16med (BxPC-3), and MUC16low (MIAPaCa-2) tumors. E and G, Serial PET-CT images of mice bearing subcutaneous S2–028 xenografts (E) and OVCAR3 xenografts (G; n = 3 mice per tumor xenograft) acquired after the intravenous administration of 2.14 mg/kg [89Zr]Zr-DFO-huAR9.6 (250 μCi; 9.25 MBq; 53 μg) showing gradual accretion of radioactivity in the tumor (T) and the liver (L) as well as gradually declining activity concentrations in the blood (indicated by the heart [H]). F and H, Longitudinal ex vivo biodistribution data acquired after the i.v. injection of 0.25 mg/kg of [89Zr]Zr-DFO-huAR9.6 (29 μCi; 1.07 MBq; 6.2 μg) in mice bearing subcutaneous S2-028 xenografts (F) and OVCAR3 xenografts (H; n = 4 mice per time point). In the graph shown in F; *, P = 0.0286. Detailed sets of %ID/g values are provided in Supplementary Tables S7 and S8.

Humanized AR9.6 demonstrates robust in vitro, in vivo, and ex vivo binding to MUC16-expressing ovarian and pancreatic cancer cells. A, Humanization of muAR9.6 by CDR grafting. B,In vitro internalization data of [89Zr]Zr-DFO-huAR9.6 in OVCAR3 cells at 1, 3, 6, and 24 hours. The blue bars represent the uptake of 10 ng of [89Zr]Zr-DFO-huAR9.6 at each timepoint, whereas the red bars represent the blocked uptake of [89Zr]Zr-DFO-huAR9.6 in the presence of a 1,000-fold excess of unlabeled huAR9.6. C,In vitro and ex vivo validation of the binding of huAR9.6 to MUC16high (OVCAR3) and MUC16neg (SKOV3) ovarian cancer cells using flow cytometry and immunohistochemical (IHC) staining of FFPE tumor sections. D,Ex vivo validation of the binding of huAR9.6 to FFPE sections from MUC16high (S2-028), MUC16med (BxPC-3), and MUC16low (MIAPaCa-2) tumors. E and G, Serial PET-CT images of mice bearing subcutaneous S2–028 xenografts (E) and OVCAR3 xenografts (G; n = 3 mice per tumor xenograft) acquired after the intravenous administration of 2.14 mg/kg [89Zr]Zr-DFO-huAR9.6 (250 μCi; 9.25 MBq; 53 μg) showing gradual accretion of radioactivity in the tumor (T) and the liver (L) as well as gradually declining activity concentrations in the blood (indicated by the heart [H]). F and H, Longitudinal ex vivo biodistribution data acquired after the i.v. injection of 0.25 mg/kg of [89Zr]Zr-DFO-huAR9.6 (29 μCi; 1.07 MBq; 6.2 μg) in mice bearing subcutaneous S2-028 xenografts (F) and OVCAR3 xenografts (H; n = 4 mice per time point). In the graph shown in F; *, P = 0.0286. Detailed sets of %ID/g values are provided in Supplementary Tables S7 and S8.