Figure 1 | Scientific Reports

Figure 1

From: An engineered tetra-valent antibody fully activates the Tie2 receptor with comparable potency to its natural ligand angiopoietin-1

Figure 1

Characterization of newly identified anti-Tie2 agonistic antibodies. (a) Cellular viability as an indicator of agonistic activity of anti-Tie2 antibodies produced by hybridoma clones 2–16 and 2–8 were assessed in human Tie2-expressing Ba/F3 compared to Ang1. Cellular viability (%) was determined using a CellTiter-Glo assay, with basal activity without any ligand (PBS) defined as 0%, and the activity of 17.9 nM (1 µg/mL) Ang1 defined as 100%. Data represent mean ± SD. (b,c) Fractionation of the antibody solution using AEX. (b) shows SDS PAGE of each fraction from AEX, and (c) shows cellular viability as an indicator of the agonistic activity of each fraction in human Tie2-expressing Ba/F3. Solid (left) arrow indicates antibody-rich fractions, dashed (right) arrow indicates the fractions with higher agonistic activity. Cellular viability (%) was determined using a CellTiter-Glo assay, with basal activity without any ligand (PBS) defined as 0%, and the activity in fraction 6 defined as 100%. (d) Cellular viability as an indicator of the agonistic activity of purified 2–16 monomer, dimer and HMW human monoclonal antibodies, isolated using SEC (Supplementary Fig. S1 and S2), in human Tie2-expressing Ba/F3 compared to Ang1. Cellular viability (%) was determined as described in (a).

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