NeuroMab™ Anti-CTNNB1 Antibody, Clone 3F9

Figure 1 shows FACS histograms of selected SIRPA antibodies binding 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. EC50 values were calculated 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 in wells with or without plate-bound CD47 protein. All CD47 blocking antibodies (1B3, 12D6, 1H11, 5F7) effectively inhibited the luminescence signal. Two CD47 non-blocking anti-SIRPA antibodies did not reduce luciferase expression. Results are expressed as background multiples. The background level is set to 1 on the y-axis.

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

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

Cells were treated with soluble full-length isotype control or soluble full-length anti-SIRPA antibody, followed by staining with DyLight650-conjugated anti-SIRPA reference antibody (SA56-DyL650) that binds to different epitope bins.

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

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

Figure 8 is a graph showing the average tumor volume of NSG mice transplanted with human immune stem cells from different cord blood donors (donors 5031, 5048, 129).

Humanized mice were implanted subcutaneously with the human breast cancer cell line MDA-MB-231, and were randomly divided into groups based on tumor volume on day -1, huCD34+ stem cell donor, body weight before randomization, and huC45+ engraftment rate before randomization. treatment group or control group. Mice were injected intraperitoneally with 40 mg/kg mouse IgG1 or 3F9 every 4 days or 10 mg/kg Keytruda every 5 days.