Fig. 5.

BsIgG 28B7/4H7 has agonistic properties as human IgG1, IgG2 and IgG4 isotype. (A) Agonistic activity of BsIgG 28B7/4H7 in human IgG1, IgG2 or IgG4 isotype backbones in GAL-ELK1 luciferase reporter assay. Luciferase activity was expressed as fold induction over an irrelevant IgG control. The data represents means ± SEM (N = 3). (B) Western blotting was conducted to test the activity of BsIgG 28B7/4H7 of human IgG1, IgG2 and IgG4 isotype on primary human adipocytes. Antibodies were used at the indicated concentrations (μg/ml) to assess a dose dependent phosphorylation of ERK (pERK). HSP90 serves as a loading control. (C) Agonistic activity of 28B7/4H7 BsIgG or an equimolar mixture of 28B7 and 4H7 over monovalent parental antibodies. Replicates of two independent experiments for each antibody are shown.