Human Anti-IL6R Recombinant Antibody (clone A7) (CAT#: HPAB-AP718-YC)

Figure 1 Generation of the dual target-directed antibody (DTA) against IL-6R and TNFα.

Binding affinities of anti-IL-6R antibody candidates (A7, B10, D2 and F2) to IL-6R. The values of dissociation constant (Kd) and coefficient of determination (R²) are displayed under the graph.

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.

Figure 2 The inhibitory effects of DTA(A7/sTNFR2) on the production of inflammatory mediators and proliferation and migration of RA-FLS.

Anti-proliferative effects of DTA(A7/TNFR2) on RA-FLS. RA-FLS were incubated with or without human TNFα, IL-6 and soluble IL-6R(sIL-6R) for 72 h. 100 μg/mL of sTNFR2, A7, DTA(A7/sTNFR2), or IgG was added during the incubation. Cell proliferation was assessed using CCK8 assay. Proliferation rates were normalized to the negative control (n.c.) value. Values are means ± SEM.

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.

Figure 3 The inhibitory effects of DTA(A7/sTNFR2) on the production of inflammatory mediators and proliferation and migration of RA-FLS.

Migration activities. Migration activities were calculated as follows: migration activity = 1—(wounded area at 16 h)/(wounded area at 0 h). The values were normalized to n.c. Mean ± SEM is presented. All results are representative of at least three independent experiments. P-values were obtained from Student's t-test (*p < 0.05; **p < 0.01; ***p < 0.001).

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.

Figure 4 Anti-osteoclastogenic activity of DTA(A7/sTNFR2).

The representative data of TRAP staining. Osteoclast precursor cells were stimulated with M-CSF and RANKL and differentiated into osteoclasts. During the stimulation, cells were incubated with IgG, TNFR2, A7, or DTA (A7/sTNFR2). Cells were fixed and stained using a TRAP staining kit after 8 days of stimulation. Scale bar: 100 μm.

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.

Figure 5 Amelioration of CIA by DTA(A7/sTNFR2).

Representative histological images of the hind paws of WT mice and CIA mice injected with IgG, sTNFR2, A7, or DTA(A7/sTNFR2). Sections were stained with H&E, safranin O, or toluidine blue. Scale bars: 400 μm in the H&E images and 200 μm in the safranin O and toluidine blue images.

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.

Figure 6 Generation of the dual target-directed antibody (DTA) against IL-6R and TNFα.

Immunoblotting of DTA candidates (A7/TNFR2, B10/TNFR2, F2/TNFR2 and D2/TNFR2) with anti-Fc antibodies in reducing and non-reducing conditions.

Kim, Y., Yi, H., Jung, H., Rim, Y. A., Park, N., Kim, J., ... & Ju, J. H. (2016). A dual target-directed agent against interleukin-6 receptor and tumor necrosis factor α ameliorates experimental arthritis. Scientific reports, 6(1), 1-12.