Journal: mAbs

Article Title: A bi-functional antibody-receptor domain fusion protein simultaneously targeting IGF-IR and VEGF for degradation

doi: 10.1080/19420862.2015.1055442

Figure Lengend Snippet: Binding to IGF-IR, VEGF and evidence for co-engagement of both targets by ELISA. ( A ) ID2 (orange), IR mAb (blue) and human IgG control (gray) binding to human IGF-IR-Fc. ( B ) ID2 (orange), FcD2 (green) and human IgG control (gray) binding to human VEGF. ( C ) ID2 (orange), IR mAb (blue) and FcD2 (green) binding to both human IGF-IR-Fc (coated) and human VEGF (detected). Each panel is a representative experiment of at least 3 repeated measurements. The graph is plotted as mean ± SEM (n = 2 ).

Article Snippet: For the VEGF-VEGFR2 blocking ELISA, the pre-incubated mixture containing 4 nM human VEGF and varied amount of antibody was added to plates coated with 40 ng/well of VEGFR2 (R&D 357-KD).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: mAbs

Article Title: A bi-functional antibody-receptor domain fusion protein simultaneously targeting IGF-IR and VEGF for degradation

doi: 10.1080/19420862.2015.1055442

Figure Lengend Snippet: Summary of ELISA binding, affinity (SPR), and ELISA blocking activities

Article Snippet: For the VEGF-VEGFR2 blocking ELISA, the pre-incubated mixture containing 4 nM human VEGF and varied amount of antibody was added to plates coated with 40 ng/well of VEGFR2 (R&D 357-KD).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Blocking Assay

Journal: mAbs

Article Title: A bi-functional antibody-receptor domain fusion protein simultaneously targeting IGF-IR and VEGF for degradation

doi: 10.1080/19420862.2015.1055442

Figure Lengend Snippet: Inhibition of VEGF-mediated endothelial cell signaling and functions by ID2. ( A ) 100 nM ID2 inhibits VEGF-induced phosphorylation of VEGFR2, downstream AKT and ERK1/2 in PAE/KDR cells as assessed by immunoblotting analysis. IR mAb and FcD2 were used as controls. ( B ) In an Oris cell migration assay, PAE/KDR cells stimulated with 100 ng/ml VEGF were treated with 100 nM ID2, IR mAb, or FcD2 for 20 hours. The fluorescence intensity of migrated cells in relative fluorescence units (RFU) was measured. ID2 significantly reduced the migration compared to VEGF and IR mAb controls ( p = 0.002 and p = 0.003, respectively, one way ANOVA). ( C ) ID2 inhibits VEGF stimulated cord formation in an ADSC/ECFC co-culture system. The total tube area for each treatment was calculated. ID2 significantly reduced the total tube area compared with VEGF only and IR mAb controls ( p < 0.0001 and p < 0.0001, respectively, one way ANOVA). ( D ) ID2 inhibits human VEGF induced HUVEC viability in a dose dependent manner in a CellTiter Glo assay. The error bar from panels B, C and D represents the SEM from each triplicate measurement.

Article Snippet: For the VEGF-VEGFR2 blocking ELISA, the pre-incubated mixture containing 4 nM human VEGF and varied amount of antibody was added to plates coated with 40 ng/well of VEGFR2 (R&D 357-KD).

Techniques: Inhibition, Western Blot, Cell Migration Assay, Fluorescence, Migration, Co-Culture Assay, Glo Assay

Journal: mAbs

Article Title: A bi-functional antibody-receptor domain fusion protein simultaneously targeting IGF-IR and VEGF for degradation

doi: 10.1080/19420862.2015.1055442

Figure Lengend Snippet: In vivo mechanism of action in Colo-205 xenograft model. ( A ) Average signal of total human IGF-IR from lysates of excised tumor, ( B ) human VEGF concentration (pg/ml) from mouse plasma and ( C ) Mouse VEGF concentration (pg/ml) from mouse plasma after 2- and 7-day treatments with saline, IR mAb, FcD2 and ID2 were determined by electrochemiluminescent assay. Significant differences in mean ( p < 0.05, n = 5) are indicated (a vs Saline; b vs IR mAb; c vs FcD2). ( D ) Percentage of nuclei with positive immunohistochemistry staining for cleaved caspase-3 from representative tumor tissue after 2-day treatment was compared and plotted as mean ± SEM (n = 5). ID2 treated group had significantly increased cleaved caspase-3 activity compared to saline control ( p = 0.0035, one way ANOVA), IR mAb ( p = 0.0041, one way ANOVA) and FcD2 ( p = 0.0254, one way ANOVA). All charts were generated and statistical analyses were performed with SigmaPlot or Graphpad Prism 6.

Article Snippet: For the VEGF-VEGFR2 blocking ELISA, the pre-incubated mixture containing 4 nM human VEGF and varied amount of antibody was added to plates coated with 40 ng/well of VEGFR2 (R&D 357-KD).

Techniques: In Vivo, Concentration Assay, Immunohistochemistry, Staining, Activity Assay, Generated