Anti-CAIX/CA9 Recombinant Antibody (Girentuximab) (CAT#: TAB-707)

Figure 1 Blood clearance of dual-labeled girentuximab over seven days after injection (p.i.) fitted to a two-compartment model.

Inset: clearance during the first three hours is depicted in more detail. Values are expressed as mean ± SD of 14 patients, since pharmacokinetic data of patient #3 were incomplete.

Hekman, M. C., Rijpkema, M., Muselaers, C. H., Oosterwijk, E., Hulsbergen-Van de Kaa, C. A., Boerman, O. C.,... & Mulders, P. F. (2018). Tumor-targeted dual-modality imaging to improve intraoperative visualization of clear cell renal cell carcinoma: a first in man study. Theranostics, 8(8), 2161.

Figure 2 Phenotypic analysis of NU12 and SK-RC-52 tumors.

Phenotypic analysis of NU12 and SK-RC-52 tumors of mice treated with sunitinib shows necrosis, decreased accumulated cG250 and decreased number of microvessels in NU12 tumors and very limited necrosis, increased accumulated cG250 and unchanged number of microvessels in SK-RC-52 tumors.A-D, NU12 control tumors; E-H, NU12 sunitinib treated tumors; I-L, SK-RC-52 control tumors; M-P, SK-RC-52 sunitinib treated tumors. HE staining in A, E, I and M. Sunitinib treatment did not modify CAIX expression in either NU12 or SK-RC-52 (B,F and J,N). In NU12 control tumors (A-D), homogeneous accumulation of cG250 (C) was observed. D: tumor vasculature visualized by staining with 9 F1. In sunitinib treated NU12 tumors (E-H), extensive necrosis was present as judged by HE (E) and accumulated cG250 (G) and microvessels (H) were only observed in the tumor
rim. SK-RC-52 control tumors (I-L), revealed focal accumulation of injected cG250 (J) and moderate microvessel density (MVD) as visualized by staining with 9 F1 (L). Accumulation of cG250 was increased in sunitinib treated SKRC52 tumors (O vs. K) and MVD appeared to be increased (P vs. L). Necrosis was limited regardless of treatment (I, M). N: necrosis. Original magnification ×25 and × 200.

Oosterwijk-Wakka, J. C., de Weijert, M. C., Franssen, G. M., Leenders, W. P., van der Laak, J. A., Boerman, O. C.,... & Oosterwijk, E. (2015). Successful combination of sunitinib and girentuximab in two renal cell carcinoma animal models: a rationale for combination treatment of patients with advanced RCC. Neoplasia, 17(2), 215-224.

Figure 3 SPECT/CT imaging of mice with SK-RC-52 tumors.

SPECT/CT analysis was performed to visualize the biodistribution and the intra-tumoral distribution of the radiolabeled antibody. Sixteen mice bearing SK-RC-52 were treated with sunitinib for 7 days and injected with 111In-Girentuximab with a specific activity of 22,5 MBq/5 μg, 3 days before start or 3 days after stop of treatment. Micro-SPECT
images of mouse bearing SK-RC-52 tumor on right flank (arrow) at 7 d after injection of 111In-girentuximab show that in addition to tumor uptake, minimal uptake in other organs was observed. More radiolabel was observed in the sunitinib treated tumors than in vehicle (A). This is in concordance with the biodistribution data. In all groups radiolabel was distributed throughout the tumor and no difference in radiolabel distribution was observed in the various treatment groups (B).

Oosterwijk-Wakka, J. C., de Weijert, M. C., Franssen, G. M., Leenders, W. P., van der Laak, J. A., Boerman, O. C.,... & Oosterwijk, E. (2015). Successful combination of sunitinib and girentuximab in two renal cell carcinoma animal models: a rationale for combination treatment of patients with advanced RCC. Neoplasia, 17(2), 215-224.

Figure 4 Immunohistochemical analyses of vital tumor specimens from different groups.

Neoadjuvant treatment with sorafenib resulted in enhanced necrosis (hematoxylin and eosin) and decreased vessel density (CD31) but had no effect on CAIX expression.

Muselaers, C. H., Stillebroer, A. B., Desar, I. M., Boers-Sonderen, M. J., van Herpen, C. M., de Weijert, M. C.,... & Mulders, P. F. (2014). Tyrosine kinase inhibitor sorafenib decreases 111In-girentuximab uptake in patients with clear cell renal cell carcinoma. Journal of Nuclear Medicine, 55(2), 242-247.

Figure 3 Anti-CAIX/CA9 Antibody (TAB-707) in Dot Blot

Dot Blot analysis of TAB-707 was performed by coating with Recombinant Human Carbonic Anhydrase IX Protein (his Tag).

TAB-707 incubation concentration: 1 ng/μL.
The secondary antibody: HRP-Anti-Human IgG (H+L)

Figure 1 Anti-CAIX/CA9 Antibody (TAB-707) in ELISA

ELISA analysis of TAB-707 was performed by coating with Recombinant Human Carbonic Anhydrase IX Protein (his Tag). Then blocked with BSA and incubated with Anti-Human DR5 Antibody (TAB-707). The HRP-conjugated goat anti-human IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 2 Anti-CAIX/CA9 Antibody (TAB-707) in WB

Western blot analysis of TAB-707 was performed by loading Recombinant Human Carbonic Anhydrase IX Protein (his Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with TAB-707 and HRP Goat Anti-Human IgG as a secondary antibody. Chemiluminescent detection was performed.