Human Anti-TYRO3 Recombinant Antibody (clone 2718) (CAT#: NS-097CN)

Figure 1 ELISA Binding assay of scFvs/IgG against hTyro 3 ECD.

A: Results for ScFvs. B: results for IgGs.

Figure 2 The purified extracellular domains of hTyro 3, hAxl or hMer were coated on micro titerplate and incubated in the presence of anti-Tyro 3 scFvs (mg/ml).

Figure 3 GAS6 competition assay with anti-Tyro3 scFvs.

Fig. A. Binding of scFvs and MAB859 to full-length Tyro3. Full-length Tyro3 was coated onto an ELISA microtiter plate. scFvs were used at 10 mg/ml and MAB859 was used at 3 concentrations: 1 mg/ml (MAB859-1), 5 mg/ml (MAB859-5) and 10 mg/ml (MAB859-10). scFv binding to full-length Tyro3 was detected with an anti-HIS-HRP antibody and MAB859 with an anti-mouse-HRP antibody. The negative controls with the detection antibodies alone were shown (anti-HIS or anti-mouse). Fig. B. Effect of scFvs on Tyro3/GAS6 binding. Biotinylated GAS6 (2 mg/ml) alone or scFv/GAS6 mixtures were applied for lh at room temperature onto full-length Tyro3 coated plates. Binding of GAS6 to Tyro3 was detected with streptavidin-HRP (400 ng/ml). The OD measured in the presence of the streptavidin-HRP alone was shown.

Figure 4 Cross-reactivity against cynomolgus Tyro3 ECD of anti-Tyro3 in the scFv (Fig. A) and IgG format (Fig. B).

Figure 5 Cross-reactivity against murine Tyro3 ECD of anti-Tyro3 in the scFv format.

Figure 6 Epitope restriction of anti-Tyro3 scFvs to structural domains of Tyro3 ECD.

Figure 7 Binding of anti-Tyro3 IgGs in living cells.

HEK293 cells transfected to express recombinant human Tyro3 and HEK293 untransfected cells were stained with the Live-Dead NIR dye and either incubated with the TYRO 3-specific MAB859 monoclonal antibody, its isotype control (upper panels) or to individual anti-Tyro3 IgG (middle and bottom panels). IgG isotype control is IGX2656. MAB002 and MAB859 binding was revealed with a PE-conjugated Goat anti-Mouse Ig Ab. Anti-Tyro3 IgG binding was revealed with a PE-conjugated Mouse-anti Human Lambda Light Chain Ab. Data were acquired using an ARIA III flow cytometer and analyzed using the Kaluza software (Beckman Coulter). Histograms shown are restricted to live cells that show a characteristic size and granulometry of HEK293 cells. Anti-Tyro3 IgG Tyro3-specific binding is shown by fluorescence intensity increase of Tyro3-transfected cells (black line) compared to untransfected cells (grey line).