BV14 binds to the membrane proximal region of VE-cadherin.

(A) Schematic representation of VE-cadherin constructs. The extracellular domains EC3 and EC4 in human VE-cadherin were substituted with the murine homologous sequence spanning from AA 215 to AA 501. Recognition binding domains of the mAbs Cad 5 and BV9 on human VE-cadherin are indicated. (B) Western blot analysis of VE-cadherin transfectant cell line extracts using anti–human and anti–mouse VE-cadherin antibodies. As expected, mAb BV13 and BV14 recognized murine VE-cadherin, while mAb Cad 5 and BV9 human VE-cadherin did not. However, only BV14 and Cad 5 could bind to the chimeric construct. (C) Fluorescence flow cytometric analysis of binding of different mAbs to endothelial cells. Saturating concentrations of mAbs were used for staining. Panels indicate fluorescence intensity, and the numbers inside the panels represent the percentage of cells positive to staining with mAbs. MAbs Cad 5 and BV9 bind human VE-cadherin cells (ii) and (iii) and BV14 binds murine VE-cadherin (iv). The analysis was performed at least 3 times for each cell type.