NeuroMab™ Anti-EPHA2 BBB Shuttle Antibody(NRZP-1022-ZP3874)

Figure 1 ITC curve of 135H11 against EphA2-LBD (Kd = 150 nM)

Figure 2 Comparison of DELFIA shift dose response curves of 123B9, 135G3 and 135H11

Figure 3 135H12 alters EphA2 cellular localization.

Figure 4 135H12 and 135H11 inhibit cell invasion. 7 x 105 BxPC3 cells were seeded into each well of a 96-well plate and processed the next day. Plates were processed once and imaged every 6 hours for 6 days.

Figure 5 135H12 and 135H11 inhibit cell migration. BxPC3 cells were plated and processed similarly to the invasion assay. Plates were imaged every 6 h for 36 h. (5C) Time-dependent inhibition of pancreatic cancer cell invasion and migration when treated with 135H11 (monomer) and 135H12 (dimer).

Figure 6 Time-dependent inhibition of pancreatic cancer cell invasion and migration when treated with 135H11 (monomer) and 135H12 (dimer). Both agonists inhibited cell invasion, while 135H12 showed a more pronounced inhibition of cell migration than its monomeric counterpart 135H11. Invasion assay n = 5, migration assay n = 10. Error bars are SEM. ****, p = 0.0001; ***, P = 0.0002; **, P = 0.003.

Figure 7 Isothermal titration calorimetry (ITC) curves of binding between sterically restricted peptides and EphA2-LBD.

Figure 8 Representative images of each treatment at time 0, 12, and 24 hours.

Figure 9 Summary of migration of BxPC3 cells over time when treated with 135H11 (monomer) and 135H12 (dimer). Error bars are SEM. ****, p=0.0001; *, P=0.01, n=4.

Figure 10A. 1D lH-aliph. 2D [15N, ¾] ultrafast HSQC spectra of 15N-EphA2-LBD NMR spectra were measured in the absence and presence of various concentrations of 135H11. Spectra were collected on a 700 MHz Bruker NMR spectrometer equipped with a TCI cryoprobe. The spectrum of the complex after titration exhibits a slow exchange in the NMR time scale.

Figure 10B. 1D lH-aliph. 2D [15N, ¾] ultrafast HSQC spectra of 15N-EphA2-LBD NMR spectra were measured in the absence and presence of various concentrations of 135H11. Spectra were collected on a 700 MHz Bruker NMR spectrometer equipped with a TCI cryoprobe. The spectrum of the complex after titration exhibits a slow exchange in the NMR time scale.