NeuroMab™ Anti-SIRPA BBB Shuttle Antibody, Clone 9C2

Figure 1 shows FACS histograms of selected SIRPA antibodies bound to rodent Chinese hamster ovary cell lines (CHO) expressing human SIRPA (HuSIRPA) or mouse SIRPA (MuSIRPA) Left panel

Figure 2 shows FACS histograms of binding of selected SIRPA antibodies to primary human macrophages.

Figure 3 shows the binding of increasing concentrations of anti-SIRPA antibodies to human SIRPA overexpressed on CHO cells. Calculate EC50 values by fitting the data to a sigmoid curve using Graph Pad Prism.

Figure 4 shows the ability of CD47-blocking and CD47-non-blocking anti-SIRPA antibodies to affect HuSIRPA-dependent luciferase expression in a cell-based reporter assay.

BWZ-HuSIRPA cells were seeded on wells with or without plate-bound CD47 protein. All CD47-blocking antibodies (1B3, 12D6, 1H11, 5F7) potently suppress luminescence signal. Two CD47-non-blocking anti-SIRPA antibodies did not reduce luciferase expression. Results are expressed as fold over background. The background level is set to 1 on y-axis.

Figure 5 shows induction of human SIRPA-dependent or human SIRPB1-dependent luciferase expression in a cell-based reporter assay.

Figure 6 shows SIRPA receptor downregulation in primary human macrophages in response to antibody stimulation.

Cells were treated with either soluble full-length isotype control or soluble full-length anti-SIRPA antibodies and subsequently stained with a DyLight650-conjugated anti-SIRPA reference antibody (SA56-DyL650) that binds to a distinct epitope bin.

Figure 7 shows enhanced phagocytic activity of macrophages treated with CD47 blocking anti-SIRPA antibodies.

Macrophages were cultured overnight in 2.5% FBS RPMI media with 5 μg/mL of 12D6, 9C5, 1H11, 5F7, 1B3, 3F9 (CD47-non-blocker) or isotype control.