Human Anti-TNFRSF10A Recombinant Antibody (clone hAY4) (CAT#: HPAB-1617-FY)

Figure 1 Characterization of bivalent and trivalent hAY4 scFvs by linkage to the LZ and ILZ domain via a hinge region, respectively.

SDS-PAGE analysis of purified hAY4 scFv antibodies. The purified proteins AY4 (∼5 g) were separated on a 12% SDS polyacrylamide gel under non-reducing conditions and visualized by staining with Coomassie Blue R-250.

Lee, S. H., Park, D. W., Sung, E. S., Park, H. R., Kim, J. K., & Kim, Y. S. (2010). Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Molecular immunology, 47(4), 816-824.

Figure 2 Characterization of the binding specificity of the humanized hAY4 scFv antibody.

Binding specificity of hAY4 and TRAIL analyzed by ELISA. ELISA plates were coated with 10 μg/ml of DR4, DR5, DcR1 and DcR2, and then incubated with 1 M of hAY4 scFv (black columns) or 50 nM of TRAIL (white columns).

Lee, S. H., Park, D. W., Sung, E. S., Park, H. R., Kim, J. K., & Kim, Y. S. (2010). Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Molecular immunology, 47(4), 816-824.

Figure 3 Comparisons of the kinetic binding interactions of hAY4, LZ-hAY4 and ILZ-hAY4 scFvs with the antigen DR4 as measured by SPR.

Representative SPR sensorgrams showing kinetic binding interactions of hAY4 with DR4.

Lee, S. H., Park, D. W., Sung, E. S., Park, H. R., Kim, J. K., & Kim, Y. S. (2010). Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Molecular immunology, 47(4), 816-824.

Figure 4 Cell killing activities of hAY4 of mAY4 IgG-sensitive cells.

Cells were incubated with the indicated concentrations (0.05–140 μg/ml) of each scFv antibody for 24 h prior to the measurement of cell viability by MTT assay.

Lee, S. H., Park, D. W., Sung, E. S., Park, H. R., Kim, J. K., & Kim, Y. S. (2010). Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Molecular immunology, 47(4), 816-824.

Figure 5 Cell death pathway induced by hAY4, LZ-hAY4 and ILZ-hAY4 scFvs.

Dual staining of Jurket cells for Annexin-V-FITC and PI following treatment with medium or 10 μg/ml of each hAY4 scFv for 20 h (as indicated in each panel). Samples were analyzed by flow cytometry.

Lee, S. H., Park, D. W., Sung, E. S., Park, H. R., Kim, J. K., & Kim, Y. S. (2010). Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Molecular immunology, 47(4), 816-824.