Anti-Human VEGFA Recombinant Antibody (Vanucizumab) (CAT#: TAB-H73)

Figure 1 Effects of vanucizumab on tumour progression.

(A) Transverse T2-weighted MR images of the central tumour slice acquired from paired mice bearing Colo205 xenografts before and 5 days after treatment, as indicated. (B) Summary of tumour volume response, determined by T2-weighted MRI, for each treatment cohort before and 5 days after treatment with vanucizumab, bevacizumab, LC06 or omalizumab (control).

Baker, L. C., Boult, J. K., Thomas, M., Koehler, A., Nayak, T., Tessier, J., ... & Horn, C. (2016). Acute tumour response to a bispecific Ang-2-VEGF-A antibody: insights from multiparametric MRI and gene expression profiling. British journal of cancer, 115(6), 691.

Figure 2 Effects of vanucizumab on tumour vascular perfusion and permeability assessed by dynamic contrast-enhanced MRI.

(A) Parametric Ktrans maps acquired from paired Colo205 xenografts before and 5 days after treatment with vanucizumab, bevacizumab, LC06 or omalizumab (control), as indicated. (B) Estimates of the transfer constant (Ktrans) and the initial area under the gadolinium uptake curve to 60 s (IAUGC60) determined from each Colo205 xenograft before and 5 days after treatment with vanucizumab, bevacizumab, LC06 or omalizumab. (C) Summary of the percentage change in Ktrans determined across all four treatment groups. There was no significant difference in the change in Ktrans 5 days post treatment with vanucizumab, bevacizumab or LC06 when compared with omalizumab (control) – two-way ANOVA with Bonferroni correction.

Baker, L. C., Boult, J. K., Thomas, M., Koehler, A., Nayak, T., Tessier, J., ... & Horn, C. (2016). Acute tumour response to a bispecific Ang-2-VEGF-A antibody: insights from multiparametric MRI and gene expression profiling. British journal of cancer, 115(6), 691.

Figure 3 Histological assessment of the effects of vanucizumab on tumour vasculature, hypoxia and necrosis.

(A) Composite fluorescence images of Hoechst 33342 uptake (blue, perfusion) and pimonidazole adduct formation (green, hypoxia) acquired from whole sections of Colo205 xenografts 5 days after treatment with vanucizumab, bevacizumab, LC06 or omalizumab (control). Fluorescence images (× 100 magnification) of CD31 immunohistochemistry used to assess tumour microvessel density (MVD) post treatment. High-magnification (× 200) images of α-SMA immunohistochemical staining used to assess pericyte coverage. Composite images of whole tumour sections stained with H&E used to assess necrosis. (B) Summary of treatment-induced changes in perfused tumour vessels, hypoxia, MVD, vascular maturation and necrosis.

Baker, L. C., Boult, J. K., Thomas, M., Koehler, A., Nayak, T., Tessier, J., ... & Horn, C. (2016). Acute tumour response to a bispecific Ang-2-VEGF-A antibody: insights from multiparametric MRI and gene expression profiling. British journal of cancer, 115(6), 691.

Figure 1 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in SDS-PAGE

SDS-PAGE analysis of TAB-H73 in non-reduced (Lane 1) and β-mercaptoethanol-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 48-63 kDa and 25 kDa, respectively.

Figure 2 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in WB

Western blot analysis of TAB-H73 was performed by loading Human VEGF165 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-H73 and HRP Goat Anti-Human IgG as a secondary antibody. Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.8 μg)

Figure 3 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in WB

Western blot analysis of TAB-H73 was performed by loading Human VEGF121 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-H73 and HRP Goat Anti-Human IgG as a secondary antibody. Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.8 μg)

Figure 4 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in DB

Dot Blot analysis of TAB-H73 was performed by coating with Human VEGF165 Protein (His Tag).

TAB-H73 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG as a secondary antibody (1: 2000)

Figure 5 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in DB

Dot Blot analysis of TAB-H73 was performed by coating with Human VEGF121 Protein (His Tag).

TAB-H73 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG as a secondary antibody (1: 2000)

Figure 6 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in ELISA

ELISA analysis of TAB-H73 was performed by coating with Human VEGF165 Protein (His Tag). Then blocked with BSA, and incubated with TAB-H73. 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 7 Anti-Human VEGFA Recombinant Antibody (TAB-H73) in ELISA

ELISA analysis of TAB-H73 was performed by coating with Human VEGF121 Protein (His Tag). Then blocked with BSA, and incubated with TAB-H73. 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.