Recombinant Humanized Antibody (AF-2) is capable of binding to IFNG, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-IFNG mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti- IFNG mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. The AF2 antibody prevented the up regulation to a greater extent than any other member of the panel.
Figure 1 Effect of AF-2, a neutralizing Ab to human IFN-γ, on PMA-induced E-selectin expression on HUVEC, pretreated with IFN-γ.
HUVEC were cultured with 200 ng/ml IFN-γ ▪ or medium alone □ for 24 h in the presence or absence of the indicated Abs at 10 μg/ml. PMA, 1 ng/ml, was then added for an additional 4 h, and cells were analyzed for expression of E-selectin by a cell-based ELISA, as described in Materials and Methods. The relative amount of E-selectin measured is indicated by the OD at 450 nm. The mean ± SD from triplicate samples in a single, representative experiment is shown. **Indicates p < 0.005; *indicates p < 0.05; NS indicates not significant, comparing with and without IFN-γ.
Melrose, J. , Tsurushita, N. , Liu, G. , & Berg, E. L. . (1998). Ifn-γ inhibits activation-induced expression of e- and p-selectin on endothelial cells. Journal of Immunology, 161(5), 2457-64.
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