Recombinant Human Anti-VEGF Antibody (Fab-12) (CAT#: PABX-185)

Figure 1 The effects of VEGF Trap, ranibizumab and bevacizumab on calcium mobilization induced byVEGF-A165 in HUVEC.

Serial dilutions of VEGF Trap (open box), ranibizumab (triangle) or bevacizumab (closed circle) were added to HUVEC along with 20 pM of VEGF-A165. The VEGF-A165 was preincubated with the inhibitors for 10 min at 25 C. The solution was added to HUVEC preloaded with fluo-4 and the fluorescence of the well was determined on a FLIPR instrument. The data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents duplicate wells at each concentration.

Papadopoulos, N., Martin, J., Ruan, Q., Rafique, A., Rosconi, M. P., Shi, E., ... & Wiegand, S. J. (2012). Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab. Angiogenesis, 15(2), 171-185.

Figure 2 Radiolabeled VEGF binding assay

[¹²⁵I]VEGF was equilibrated (23 °C) with serial dilutions of unlabeled VEGF and (a) Fab-12 or (c) Y0317. Fabs were captured with an anti-Fab antibody-coated immunosorbant plate. Scatchard analysis (Munson & Rodbard, 1980) with a 1:1 binding model was used to calculate Kd of (b) 433 (116) pM for Fab-12 and (d) 19.8(4.3) pM for Y0317.

Clark, L. A., Boriack-Sjodin, P. A., Day, E., Eldredge, J., Fitch, C., Jarpe, M., ... & Van Vlijmen, H. W. (2008). An antibody loop replacement design feasibility study and a loop-swapped dimer structure. Protein Engineering, Design & Selection, 22(2), 93-101.