Anti-Human FLT3 Recombinant Antibody (IMC-EB10) (CAT#: TAB-291CL)

Figure 1 ALL derived cell lines expressing wt FLT3 or FLT3-D835H (on Hb1119 cells) were treated with 10 μg/mL IMC-EB10 for 1 hour.

Immunoprecipitates and total protein extracts were resolved by 8% or 10% SDS-PAGE, respectively, and subjected to immunoblot analysis with the indicated phospho-specific antibodies. The same blots were then stripped and reprobed with protein-specific antibodies.

Piloto, O., Nguyen, B., Huso, D., Kim, K. T., Li, Y., Witte, L., ... & Small, D. (2006). IMC-EB10, an anti-FLT3 monoclonal antibody, prolongs survival and reduces nonobese diabetic/severe combined immunodeficient engraftment of some acute lymphoblastic leukemia cell lines and primary leukemic samples. Cancer research, 66(9), 4843-4851.

Figure 2 IMC-EB10 significantly reduces engraftment of primary ALL blasts in NOD/SCID mice.

NOD/SCID mice received 1×10⁶ primary ALL blast via tail vein injection and were treated with 400 Ag IMC-EB10 i.p. thrice weekly starting 24 hours after cell injection. Control mice received 200 AL PBS or 400 Ag IMC-C225 i.p. thrice weekly starting 24 hours after cell injection. Bone marrow sections of mice were stained with H&E for morphology.

Piloto, O., Nguyen, B., Huso, D., Kim, K. T., Li, Y., Witte, L., ... & Small, D. (2006). IMC-EB10, an anti-FLT3 monoclonal antibody, prolongs survival and reduces nonobese diabetic/severe combined immunodeficient engraftment of some acute lymphoblastic leukemia cell lines and primary leukemic samples. Cancer research, 66(9), 4843-4851.

Figure 3 IMC-EB10 can inhibit or activate FLT3 phosphorylation along with downstream STAT5, Akt, and MAPK signaling.

ALL-derived cell lines were stained with CD135-PE or isotype control and analyzed by FACS.

Piloto, O., Nguyen, B., Huso, D., Kim, K. T., Li, Y., Witte, L., ... & Small, D. (2006). IMC-EB10, an anti-FLT3 monoclonal antibody, prolongs survival and reduces nonobese diabetic/severe combined immunodeficient engraftment of some acute lymphoblastic leukemia cell lines and primary leukemic samples. Cancer research, 66(9), 4843-4851.

Figure 4 IMC-EB10 prolongs survival of NOD/SCID mice given SEM-K2 cells.

NOD/SCID mice given 0.5×10⁶ SEM-K2 cells via tail vein injection were injected i.p. with 400 μg IMC-C225 thrice weekly starting 24 hours after cell injection as control treatment or 400 μg IMC-EB10 thrice weekly starting 24 hours after cell injection, once starting 24 hours after cell injection, thrice every other day
starting 24 hours after cell injection, or thrice weekly starting 7 days after cell injection. Mice were monitored daily for survival.

Piloto, O., Nguyen, B., Huso, D., Kim, K. T., Li, Y., Witte, L., ... & Small, D. (2006). IMC-EB10, an anti-FLT3 monoclonal antibody, prolongs survival and reduces nonobese diabetic/severe combined immunodeficient engraftment of some acute lymphoblastic leukemia cell lines and primary leukemic samples. Cancer research, 66(9), 4843-4851.

Figure 5 EB10 bind to recombinant FLT3-Fc protein and FLT3 expressed on leukemia cells.

FLT3-Fc (His) protein was immobilized onto 96-well-plates precoated with an anti-Fc antibody. Serial dilutions of EB10 was added and incubated for 1 h, followed by incubation with an antihuman Fab-HRP conjugate.

Williams, B., Atkins, A., Zhang, H., Lu, D., Jimenez, X., Li, H., ... & Li, Y. (2005). Cell-based selection of internalizing fully human antagonistic antibodies directed against FLT3 for suppression of leukemia cell growth. Leukemia, 19(8), 1432.

Figure 6 Binding of EB10 to cell surface-expressed FLT3.

FLT3-negative JM1 cells were incubated with EB10, followed by a PE-conjugated antihuman κ antibody, and analyzed using a flow cytometer.

Williams, B., Atkins, A., Zhang, H., Lu, D., Jimenez, X., Li, H., ... & Li, Y. (2005). Cell-based selection of internalizing fully human antagonistic antibodies directed against FLT3 for suppression of leukemia cell growth. Leukemia, 19(8), 1432.

Figure 1 Anti-Human FLT3 Therapeutic Antibody (TAB-291CL) in DB

Dot Blot analysis of TAB-291CL was performed by coating with human FLT3 protein (His tag).

TAB-291CL incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG was used as a secondary antibody (1: 6000)

Figure 2 Anti-Human FLT3 Therapeutic Antibody (TAB-291CL) in WB

Western blot analysis of TAB-291CL was performed with human FLT3 protein onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with TAB-291CL and HRP goat anti-Human IgG was used as a secondary antibody (1: 6000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.2 μg)
Lane 2: Reduced antigen (0.4 μg)
Lane 3: Reduced antigen (0.8 μg)

Figure 3 Anti-Human FLT3 Therapeutic Antibody (TAB-291CL) in ELISA

ELISA analysis of TAB-291CL was performed by coating with human FLT3 protein (His tag). Then blocked with BSA and incubated with TAB-291CL. The HRP-conjugated goat anti-Human IgG was used as a secondary antibody (1: 6000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.