Afuco™ Anti-Human PDGFRB ADCC Recombinant Antibody (IMC-2C5), ADCC Enhanced (CAT#: AFC-678CL)

Figure 1 IMC-2C5 binds to human and mouse PDGFRβ and efficiently blocks PDGF-B/PDGFRβ interaction.

1B3, an antibody specific to mouse PDGFRβ, was used as a control. The binding activities of IMC-2C5 to immobilized human and mouse PDGFRβ were assayed by ELISA. (A) Binding to hPDGFRβ. (B) Binding to mPDGFRβ. (C) Blocking PDGF-B binding to hPDGFRβ. (D) Blocking PDGF-B binding to mPDGFRβ.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.

Figure 2 IMC-2C5 antibody inhibits PDGF-B-stimulated receptor phosphorylation and downstream signaling.

Caki-1 (human), NIH/3T3, and D122 (mouse) cells were treated as described in the Materials and Methods section. The receptors were immunoprecipitated from the cell lysate supernatant by anti-PDGFRβ antibodies and were resolved by SDS-PAGE. The amount of phosphorylated PDGFRβ (p-PDGFRβ) was assessed through immunoblot analysis using an anti-phospho-tyrosine antibody-HRP conjugate. The amounts of total PDGFRβ proteins loaded in each lane were confirmed by detecting with antibodies to PDGFRβ. To analyze the effect of IMC-2C5 on downstream signaling, the cell lysates were resolved by SDS-PAGE. Phosphorylated Akt and p44/42 MAP kinase, as well as total loadings of Akt and p44/42 MAP kinase, were then detected using Western blot analysis. (A) Caki-1. (B) NIH/3T3. (C) D122.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.

Figure 3 IMC-2C5 antibody inhibits tumor growth as a monotherapy agent in xenograft models.

nu/nu mice (female, 7-8 weeks) were injected subcutaneously with ∼5 x 106 tumor cells: OVCAR-8 (A), NCI-H460 (B), OVCAR-5 (C), or Caki-1 (D). When tumors reached 250 to 300 mm3, mice were randomized into two treatment groups (n = 10 or 12) following a treatment schedule: antibodies at 60 mg/kg twice weekly (A, C, and D) or three times a week at 40 mg/kg (B) by i.p. injection. Human IgG and USP saline were used as controls. Tumor volumes of mice were recorded twice a week. Results shown are the mean ± SEM.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.

Figure 4 IMC-2C5 enhances the antitumor activity of an anti-VEGFR2 antibody, DC101.

Athymic nude mice bearing xenografted tumors of 200 to 300 mm3 size were randomized and divided into four treatment groups (n = 12 per group) and were treated by i.p. injections three times a week with USP saline, IMC-2C5 (40 mg/kg), DC101 (40 mg/kg), or IMC-2C5 plus DC101 (40 mg/kg each). (A) BxPC-3. (B) NCI-H460. (C) MIA-PaCa-2. (D) NCI-H292. (E) Detroit-562. (F) HCT-116.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.

Figure 5 IMC-2C5 affects the levels of expression of several tumor-associated growth factors in the antibody treated tumors.

NCI-H460 cell suspension (5 x 106 cells per mouse), was subcutaneously injected into female athymic mice. When tumors reached ∼250 mm3, mice were randomized and divided into four treatment groups and were treated by i.p. injection three times a week with USP saline, IMC-2C5 (40 mg/kg), DC101 (40 mg/kg), or IMC-2C5 plus DC101 (40 mg/kg each). Six tumors from each treatment group were harvested at days 3, 7, and 14 and were homogenized. ELISA was performed to estimate the levels of PDGF-B (A), VEGF (B), HIF-1α (C), and bFGF (D) in each treated tumors.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.

Figure 6 IMC-2C5 has an additive effect to DC101/chemotherapy in xenograft models.

Athymic nude mice bearing xenografted tumors of 350 to 400 mm3 size were randomized and divided into four treatment groups (n = 12 per group) and were treated by i.p. injections three times a week with USP saline, DC101 (40 mg/kg), or IMC-2C5 plus DC101 (40 mg/kg each). Chemotherapy drugs were dosed i.p. once a week, which started 1 day before the antibody treatment as indicated. (A) NCI-H460 and paclitaxel at 20 mg/kg. (B) MIA-PaCa-2 and gemcitabine at 500 mg/kg.

Shen, J., Vil, M. D., Prewett, M., Damoci, C., Zhang, H., Li, H.,... & Bassi, R. (2009). Development of a fully human anti-PDGFRβ antibody that suppresses growth of human tumor xenografts and enhances antitumor activity of an anti-VEGFR2 antibody. Neoplasia, 11(6), 594-604.