Anti-Human EGFR Recombinant Antibody (Panitumumab) (CAT#: TAB-020)

Figure 1 EGFR G465R mutant reveals almost complete abrogation of panitumumab and cetuximab binding.

EGFR mutation G465R abrogates binding of panitumumab and cetuximab at the protein level. Wile-type and mutant EGFR-Fc proteins were expressed and the binding of therapeutic antibodies to immobilized proteins was assessed by ELISA. Data are means from 3 experiments +/- SEM. * p<0.05, ** p<0.01, n.s. = not significant (student's T-test comparing binding of respective antibody to mutant versus wt EGFR-Fc).

Braig, F., März, M., Schieferdecker, A., Schulte, A., Voigt, M., Stein, A., ... & Engel, E. (2015). Epidermal growth factor receptor mutation mediates cross-resistance to panitumumab and cetuximab in gastrointestinal cancer. Oncotarget, 6(14), 12035.

Figure 2 EGFR G465R mutant reveals almost complete abrogation of panitumumab and cetuximab binding.

Panitumumab and cetuximab binding is abrogated in CHO cells transfected with EGFR G465R . EGFR negative CHO cells were transfected with wild type EGFR or mutants thereof. Binding of panitumumab, cetuximab or a control polyclonal EGFR antibody was assessed by FACS analysis 48h after transfection. FSC = forward scatter.

Braig, F., März, M., Schieferdecker, A., Schulte, A., Voigt, M., Stein, A., ... & Engel, E. (2015). Epidermal growth factor receptor mutation mediates cross-resistance to panitumumab and cetuximab in gastrointestinal cancer. Oncotarget, 6(14), 12035.

Figure 3 EGFR G465R mutation induces cross-resistance to panitumumab and cetuximab in an EGF-dependent Ba/ F3 cellular model.

A: EGFR signaling in EGF-dependent Ba/F3 model. Wt and S492R or G465R mutant EGFR-expressing Ba/F3 cells were cultured in the presence or absence of EGF and with addition of cetuximab, panitumumab, rituximab or erlotinib. After 2 hours, cells were harvested and EGFR/pEGFR expression analyzed by western blot analysis. B: Sensitivity of EGFR wt or EGFR G465R mutant- transfected Ba/F3 cells to treatment with EGFR-targeted antibodies. Ba/F3 cells were transformed to IL-3 independence with EGFR wt or mutant constructs and subsequently cultured in the presence or absence of EGF or with EGF in combination with panitumumab, cetuximab or control antibody rituximab. The number of viable cells was determined by trypan blue exclusion every 12 hours beginning 24 hours after seeding and plotted. Data are means from triplicate experiments +/-SEM.

Braig, F., März, M., Schieferdecker, A., Schulte, A., Voigt, M., Stein, A., ... & Engel, E. (2015). Epidermal growth factor receptor mutation mediates cross-resistance to panitumumab and cetuximab in gastrointestinal cancer. Oncotarget, 6(14), 12035.

Figure 4 In vitro EGFR targeting of panitumumab.

Immunofluorescence staining of UM-SCC-22B tumor cells and tissues for EGFR using panitumumab as the primary antibody.

Liu, Z., Liu, Y., Jia, B., Zhao, H., Jin, X., Li, F., ... & Wang, F. (2010). Epidermal growth factor receptor–targeted radioimmunotherapy of human head and neck cancer xenografts using 90Y-labeled fully human antibody panitumumab. Molecular cancer therapeutics, 1535-7163.

Figure 5 In vitro EGFR targeting of panitumumab.

In vitro inhibition of 125I-panitumumab binding to EGFR on UM-SCC-22B human head and neck cancer cells by DOTA-panitumumab and panitumumab. Points, mean (n = 3); bars, SD.

Liu, Z., Liu, Y., Jia, B., Zhao, H., Jin, X., Li, F., ... & Wang, F. (2010). Epidermal growth factor receptor–targeted radioimmunotherapy of human head and neck cancer xenografts using 90Y-labeled fully human antibody panitumumab. Molecular cancer therapeutics, 1535-7163.

Figure 1 Anti-Human EGFR Recombinant Antibody (TAB-020) in WB

Western blot analysis of TAB-020 was performed with Human EGFR protein.

TAB-020 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-IgG (H+L)
Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.3 μg)
Lane 3: Reducing Antigen (0.6 μg)

Figure 2 Anti-Human EGFR Recombinant Antibody (TAB-020) in ELISA

ELISA analysis of TAB-020 was performed by coating with Human EGFR protein.

The secondary antibody: HRP-anti-Human IgG