Human Anti-TNFRSF12A Recombinant Antibody (clone TPP-2090) (CAT#: HPAB-0049-LSX)

Figure 1 Interaction of TWEAKR ectodomain with antibodies and reference antibodies.

Shown is the result of an ELISA with TWEAKR-Fc fusion protein (TPP-2202) coating (1 mg/ml) and 0.08 mg/ml (open bars) and 0.3 mg/ml (filled bars) of biotinylated IgG as soluble binding partner. Detection was done with Streptavidin-HRP and Amplex-Red substrate. Y is "ELISA signal intensity [Rfu]"; X are "antibody constructs tested": a is "TPP-2090"; b is "TPP-2084"; c is "PDL-192(TPP-1104)"; d is "P4A8(TPP-1324)"; e is "P3G5(TPP-2195)"; f is "136.1(TPP-2194)"; h is "ITEM1"; i is "ITEM4"; j is a murine isotype control; k is a human isotype control. All tested antibodies show saturated binding with a concentration of 80 ng/ml.

Figure 2 Interaction of TWEAKR cysteine rich domain with antibodies and reference antibodies.

Shown is the result of an ELISA with TWEAKR(34-68)-Fc fusion protein (TPP-2203) coating (1 mg/ml) and 0.08 mg/ml (open bars) and 0.3 mg/ml (filled bars) of biotinylated IgG as soluble binding partner. Detection was done with Streptavidin-HRP and Amplex-Red substrate. Y is "ELISA signal intensity [Rfu]"; X are "antibody constructs tested": a is "TPP-2090"; b is "TPP-2084"; c is "PDL-192(TPP-1104)"; d is "P4A8(TPP-1324)"; e is "P3G5(TPP-2195)"; f is "136.1(TPP-2194)"; h is "ITEM1"; i is "ITEM4"; j is a murine isotype control; k is a human isotype control. The antibodies bind to the cysteine rich domain.

Figure 3 Induction of Stat-1 and NF-kappaB2 signaling pathways by anti-TWEAKR antibodies in vivo.

Treatment of mice started at day 7 after tumor cell inoculation and lysates were prepared from snap frozen tumors taken at the end of the study (day 29). Blots from 2 representative animals per group are shown. Treatment with TPP-2090 leads to a strong induction of P-Stat 1 & Total Stat 1 levels as well as NF-kappaB2 activation (shown by the appearance of the p52 band) in WiDr xenograft tumors. Lysates of snap frozen WiDr xenograft tumors after treatment with PBS (i.v., q4dx7: lanes 1&2) or TPP-2090 (3 mg/kg, i.v., q4dx7: lanes 3&4) were subjected to Western Blot analysis detected with specific antibodies for P-Stat 1 (a), Stat-1 (b), NF-kappa2-p52 (c) and GAPDH (d).

Figure 4 Efficacy of 10 mg/kg TPP-2090 (i.v., q4dx8) in the human lung cancer xenograft NCI-H322 in monotherapy and combination therapy with Paclitaxel (16 mg/kg, i.v., q7dx4).

Treatment started 14d after inoculation with established tumors of about 45 mm2. A is "Vehicle group, treated with PBS (i.v. q4dx8)". B is "TPP-2090, 5 mg/kg", C is "TPP-2090, 10 mg/kg", D is "Paclitaxel, 16 mg/kg", E is "Combo TPP-2090 10 mg/kg+Paclitaxel 16 mg/kg". (X is "Time after inoculation [days]"; Y is "Tumor area, means of n=10; SD [mm²]")

Figure 5 Efficacy of anti-TWEAKR antibodies in the human renal cell cancer xenograft 786-O after treatment with 0.3, 1.0 and 3.0 mg/kg (i.v., q4dx3) started at day 7 after tumor cell inoculation.

Shown are final tumor weights at day 40. A is "Vehicle group, treated with PBS (i.v. q4dx3)". B is "Isotype, 3 mg/kg", C is "TPP-2084, 0.3 mg/kg", D is "TPP-2084, 1 mg/kg", E is "TPP-2084, 3 mg/kg", F is "TPP-2090, 0.3 mg/kg", G is "TPP-2090, 1 mg/kg", H is "TPP-2090, 3 mg/kg". (Y is "Tumor weights means of n=8; SD [g]").