Afuco™ Anti-Human IGF2 ADCC Recombinant Antibody (DX-2647), ADCC Enhanced (CAT#: AFC-372CL)

Figure 1 DX-2647 blocks IGF-II-stimulated proliferation.

A, IGF-II/IGF-IIE synergizes with IL-4 to stimulate Ba/F3 cell proliferation. Ba/F3 cells were stimulated with the indicated cytokines, growth factors, or combinations thereof in the presence or absence of lipopolysaccharide (LPS). Proliferation was assessed after 2 d. B, representative plot of DX-2647-induced inhibition of IGF-II-stimulated and IGF-IIE (100 ng/mL)-stimulated Ba/F3 cell proliferation. n = 3 for each experiment.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.

Figure 2 DX-2647 binds to IGF-II with high affinity.

A, kinetic analysis of IGF-II, IGF-IIE, and IGF-I binding to DX-2647. Mean values and SDs for the kinetic parameters Ka and Kd were determined from n independent experiments as 1.8 ± 0.7 × 10⁶ mol/L⁻¹ s⁻¹ and 8.9 ± 4.8 × 10⁻⁵ s⁻¹ for IGF-II (n = 4), 1.9 ± 1.3 × 10⁶ mol/L⁻¹ s⁻¹ and 2.0 ± 1.0 10⁻⁵ s⁻¹ for IGF-IIE (n = 4), and 2.5 ± 0.9 × 10⁶ mol/L⁻¹ s⁻¹ and 2.4 ± 0.4 10⁻² s⁻¹ for IGF-I. Average KD values for IGF-II, IGF-IIE, and IGF-I were calculated as 49.4 pmol/L, 10.5 pmol/L, and 9.6 nmol/L, respectively. B, representative SPR tracing and tabular data showing that DX-2647 does not bind to precomplexed IGF-II.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.

Figure 3 DX-2647 blocks IGF-driven cell signaling events.

Dose-dependent inhibition of IGF-II stimulated phosphorylation of IGF-IR (A) or IR-A (B) in transfected NIH-3T3 cells. C, DX-2647 blocks exogenous and endogenous IGF-II/IGF-I-mediated phosphorylation of IGF-IR in select cancer cell lines. n = 3 for each experiment.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.

Figure 4 DX-2647 inhibits anchorage-independent and anchorage-dependent colony formation in HCC cell lines.

A, HepG2 cell anchorage-independent colony formation. Representative images of colonies are shown (inset). B, Hep3B-cell anchorage-dependent colony formation. Representative images for each respective dose of DX-2647 tested are shown above each bar. n = 3 for each experiment.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.

Figure 5 DX-2647 slows tumor progression in the Hep3B hepatocellular xenograft model.

A, tumor burden graph depicting slowing of tumor progression by DX-2647. B, tumor wet weight after tumor excision (day 39). C, animal body weight over time indicates no significant changes due to treatment regimen. Number of animals for each group is eight to nine. *, P < 0.05, compared with PBS-treated control group.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.

Figure 6 Immunohistochemical analysis of tumors from DX-2647-treated animals reveals decreased CD31 staining and reduced levels of IGF-IIE and IGF-IR phosphorylation levels.

A, H&E and immunohistochemical evaluation of human xenograft Hep3B tumors. Tumor sections were stained with H&E and immunohistochemistry for human IgG1 (hIgG1), TUNEL assay (brown), and CD31 (red) as described in Materials and Methods. Pictures are representative of four animals from each group. Apo, apoptosis. Dotted line delineates apoptotic areas. Magnifications, ×400 (a, b, c1-c3, and g-o) and ×100 (d-f). B, immunohistochemical evaluation IGF pathway molecules. Tumor sections were stained for IGF-IIE, IGF-II, IGF-I, IGF-IR, and IGF-IR-phospho as described in Materials and Methods. Pictures are representative of four animals from each group.

Dransfield, D. T., Cohen, E. H., Chang, Q., Sparrow, L. G., Bentley, J. D., Dolezal, O., ... & Phan, T. (2010). A human monoclonal antibody against insulin-like growth factor-II blocks the growth of human hepatocellular carcinoma cell lines in vitro and in vivo. Molecular cancer therapeutics, 9(6), 1809-1819.