NeuroMab™ Anti-CD284 Antibody(NRP-0422-P739)

Figure 1 is a graph depicting the binding of a murine monoclonal antibody (referred to herein as "18H10") to the TLR4/MD-2 complex.

Binding specificity was shown by flow cytometry using mock-transfected or TLR4/MD-2-transfected cells. Results for mock-transfected cells are shown in the filled plot (left), while results for cells transfected with TLR4/MD-2 are shown in the outline plot (right).

Figure 2 is a graph depicting inhibition of lipopolysaccharide (LPS)-induced IL-8 production in TLR4/MD-2 transfected HEK 293 cells by monoclonal antibody mu18H10.

Cells were incubated with indicated concentrations of mu18H10, HTA 125 (commercially available anti-human TLR4 non-blocking monoclonal antibody) or antibody control followed by incubation with LPS (15 ng/ml). IL-8 levels were assessed 16 hours after LPS treatment.

Figure 3 depicts the inhibition of LPS-induced IL-8 production in human whole blood by monoclonal antibody mu18H10.

Cells were incubated with indicated concentrations of mu18H10, HTA 125 (commercially available anti-human TLR4 non-blocking monoclonal antibody) or antibody control followed by incubation with LPS (15 ng/ml). IL-8 levels were assessed 16 hours after LPS treatment.

Figure 4 is a graph demonstrating the lack of specificity of mu18H10 for recombinant soluble MD-2 purified from supernatants of baculovirus-infected insect cells, as determined by ELISA.

Cells were incubated with indicated concentrations of mu18H10, HTA 125 (commercially available anti-human TLR4 non-blocking monoclonal antibody) or antibody control followed by incubation with LPS (15 ng/ml). IL-8 levels were assessed 16 hours after LPS treatment.

Figure 5 is a graph depicting the ability of the VH and VL nucleotide sequences of mu18H10 expressed as a chimeric MAb ("chimeric 18H10") to specifically bind human TLR4/MD-2 complexes on the surface of transfected CHO cells.

Cells were incubated with indicated concentrations of mu18H10, HTA 125 (commercially available anti-human TLR4 non-blocking monoclonal antibody) or antibody control followed by incubation with LPS (15 ng/ml). IL-8 levels were assessed 16 hours after LPS treatment.

Figure 6 is a graph depicting inhibition of lipopolysaccharide (LPS)-induced IL-8 production by chimeric 18H10 MAbs in TLR4/MD-2 transfected HEK 293 cells.

Cells were incubated with indicated concentrations of mu18H10, HTA 125 (commercially available anti-human TLR4 non-blocking monoclonal antibody) or antibody control followed by incubation with LPS (15 ng/ml). IL-8 levels were assessed 16 hours after LPS treatment.