We generated an advanced version of ABTAA, which recognizes the different epitope of human ANGPT2 and has a higher production rate in the mammalian cells. The properties of the new version of ABTAA were almost similar to the original version of ABTAA (19). The new version binds to human ANGPT2 with an equilibrium dissociation constant (Kd) value of 0.9 nM (fig. S10A). Furthermore, in the presence of ANGPT2 in primarily cultured human umbilical vein ECs (HUVECs; Lonza), the ABTAA triggered TIE2 phosphorylation and activation of its downstream signals, including Akt and extracellular signal–regulated kinase (ERK), in a dose-dependent manner (fig. S10B). ABTAA + ANGPT2 also induced TIE2 translocation to cell-cell junction and nuclear clearance of FOXO1 in common with ANGPT1 (fig. S10, C and D).

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