Anti-Human CCR4 Recombinant Antibody (308) (CAT#: TAB-1523CL)

Figure 1 Binding of anti-CCR4 scFvs to target cells, expressing CCR4.

ScFvs were cross-linked using anti-Myc-antibody (mouse) to simulate a dimeric situation and diluted 3-fold, starting at 10 μg/mL. Bound scFvs were detected using RPE-conjugated anti-mouse IgG. a.) Binding to CCR4-positive DT40 cells, b.) Binding to CCR4-negative DT40 cells.

Figure 2 Binding of anti-CCR4 scFvs to natural target cell (CCRF-CEM), expressing CCR4.

ScFvs were cross-linked using anti-Myc-antibody (mouse) to simulate a dimeric situation and diluted 3-fold, starting at 20 μg/mL. Bound scFvs were detected using RPE-conjugated anti-mouse IgG.

Figure 3 Ligand-interfering binding experiments of anti-CCR4 scFvs in presence of TARC and MDC.

ScFvs were incubated at a concentration of 0.5 μg/mL in presence of 2.5 μg/mL mouse anti-Myc-antibody (mouse) in presence of a fixed concentration of ligands (1 μg/mL) and stained using an anti-mouse-RPE conjugated antibody. Signals were compared to cells, stained only with antibody-controls (background) as well as to unstained cells (unstained).

Figure 4 Competition experiment between anti-CCR4 antibodies 208, 306, 308, 406, 501, 503, 601, 60, 803 and KM3060var in presence of biotinylated KW0761-lgG.

The biotinylated KW0761-lgG was detected using PE-conjugated Streptavidin.

Figure 5 Inhibition of TARC-mediated Ca-Flux by anti-CCR4 scFv antibodies in comparison to KM3060var.

TARC was incubated at a final concentration of 28.6 ng/μL (3.6 nM) in presence of increasing scFv-concentrations, titrated over six dilution points, starting at 0.5 μg/mL. The signals were expressed in % where 100% signaling refers to recorded signals in presence of TARC-ligand, but no scFv antibody.

Figure 6 Inhibition of MDC-mediated Ca-Flux by anti-CCR4 scFv antibodies.

MDC was incubated at a final concentration of 5 ng/μL (6.25 nM) in presence of a fixed scFv concentration of 10 μg/mL. The signals were expressed in % where 100% signaling refers to recorded signals in presence of MDC-ligand, but no scFv antibody.

Figure 7 Inhibition of TARC-mediated chemotaxis on CCR4+ CCRF-CEM cells in presence of increasing concentrations of anti-CCR4 scFv antibodies (0.36 to 333 nM).

TARC had a fixed concentration of 3.5 nM. The signals were expressed in % where 100% refers to migration of cells in presence of TARC-ligand, but no scFv antibody. The graphs were fitted.