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

Figure 1.

Figure 1. In vitro and in vivo characterization of the binding of muAR9.6 to ovarian cancer cells. A and B, Flow cytometry illustrating the strong binding of muAR9.6 to OVCAR3 cells as well as its marginal binding to SKOV3 cells. C, RBA showing the high (81.4% ± 2.1%) and blockable (11% ± 1%) binding of [89Zr]Zr-DFO-muAR9.6 to MUC16-positive OVCAR3 cells as well as its low (15.2% ± 1.3%) binding to MUC16-negative SKOV3 cells. The inset shows cellular internalization of [89Zr]Zr-DFO-muAR9.6 between 1 hour and 24 hours after adding to ∼1 million OVCAR3 cells. The blue bars represent the uptake of 10 ng of [89Zr]Zr-DFO-muAR9.6 at each timepoint, whereas the red bars represent the blocked uptake of [89Zr]Zr-DFO-muAR9.6 in the presence of a 1,000-fold excess of unlabeled muAR9.6. D, Immunostaining and fluorescence microscopy of OVCAR3 cells with muAR9.6 and FITC-labeled muAR9.6 revealing the membrane-bound localization of fluorescence. E, PET images acquired 5 days after the administration of the 1.2 ± 0.1 mg/kg [89Zr]Zr-DFO-muAR9.6 (255 ± 49.5 μCi; 9.4 ± 1.8 MBq; 29.6 ± 2.0 μg) in female Nu/Nu mice bearing OVCAR3 and SKOV3 subcutaneous xenografts (n = 3 mice per tumor type). The differential uptake of the radioimmunoconjugate in the tumors (T) can be seen as well as accumulation in other tissue compartments, including the heart [H], liver [L], and bone [B]. The latter is the result of the accretion of free, osteophilic [89Zr]Zr4+ released from the radioimmunoconjugate. Serial PET images are shown in Supplementary Figs. S7 and S8. F, Ex vivo biodistribution profile up to 5 days after the intravenous administration of 0.11 ± 0.02 mg/kg of [89Zr]Zr-DFO-muAR9.6 (24 ± 1.4 μCi; 0.88 ± 0.05 MBq; 2.8 ± 0.5 μg) to female Nu/Nu mice bearing OVCAR3 and SKOV3 subcutaneous xenografts (n = 5 mice per tumor type). *, P ≤ 0.03; **, P ≤ 0.01; ***, P ≤ 0.0005. Detailed sets of %ID/g values are provided in Supplementary Tables S1 and S2. The maximum intensity projections have been scaled from 0% to 100%.

In vitro and in vivo characterization of the binding of muAR9.6 to ovarian cancer cells. A and B, Flow cytometry illustrating the strong binding of muAR9.6 to OVCAR3 cells as well as its marginal binding to SKOV3 cells. C, RBA showing the high (81.4% ± 2.1%) and blockable (11% ± 1%) binding of [89Zr]Zr-DFO-muAR9.6 to MUC16-positive OVCAR3 cells as well as its low (15.2% ± 1.3%) binding to MUC16-negative SKOV3 cells. The inset shows cellular internalization of [89Zr]Zr-DFO-muAR9.6 between 1 hour and 24 hours after adding to ∼1 million OVCAR3 cells. The blue bars represent the uptake of 10 ng of [89Zr]Zr-DFO-muAR9.6 at each timepoint, whereas the red bars represent the blocked uptake of [89Zr]Zr-DFO-muAR9.6 in the presence of a 1,000-fold excess of unlabeled muAR9.6. D, Immunostaining and fluorescence microscopy of OVCAR3 cells with muAR9.6 and FITC-labeled muAR9.6 revealing the membrane-bound localization of fluorescence. E, PET images acquired 5 days after the administration of the 1.2 ± 0.1 mg/kg [89Zr]Zr-DFO-muAR9.6 (255 ± 49.5 μCi; 9.4 ± 1.8 MBq; 29.6 ± 2.0 μg) in female Nu/Nu mice bearing OVCAR3 and SKOV3 subcutaneous xenografts (n = 3 mice per tumor type). The differential uptake of the radioimmunoconjugate in the tumors (T) can be seen as well as accumulation in other tissue compartments, including the heart [H], liver [L], and bone [B]. The latter is the result of the accretion of free, osteophilic [89Zr]Zr4+ released from the radioimmunoconjugate. Serial PET images are shown in Supplementary Figs. S7 and S8. F,Ex vivo biodistribution profile up to 5 days after the intravenous administration of 0.11 ± 0.02 mg/kg of [89Zr]Zr-DFO-muAR9.6 (24 ± 1.4 μCi; 0.88 ± 0.05 MBq; 2.8 ± 0.5 μg) to female Nu/Nu mice bearing OVCAR3 and SKOV3 subcutaneous xenografts (n = 5 mice per tumor type). *, P ≤ 0.03; **, P ≤ 0.01; ***, P ≤ 0.0005. Detailed sets of %ID/g values are provided in Supplementary Tables S1 and S2. The maximum intensity projections have been scaled from 0% to 100%.