Anti-CD33 Recombinant Antibody (Lintuzumab) (CAT#: TAB-756)

Figure 1 Reductions in bone marrow blasts were seen at all dose levels.

Percentage change in bone marrow blasts after treatment with sequential cytarabine and 213 Bi-lintuzumab in 26 evaluable patients.

Rosenblat, T. L., McDevitt, M. R., Mulford, D. A., Pandit-Taskar, N., Divgi, C. R., Panageas, K. S., ... & Larson, S. M. (2010). Sequential cytarabine and α-particle immunotherapy with bismuth-213–lintuzumab (HuM195) for acute myeloid leukemia. Clinical Cancer Research, 1078-0432.

Figure 2 Flow cytometry-based cytotoxicity assay for quantification of AMG330-specific lysis of myeloid cells.

The CD33+ cell line U937 was co-incubated for 24 h with MNC from a healthy donor at an E:T ratio of 1:1 (referred to CD3+ T-cell) in the presence of AMG330 or Control Ab constructs. Dot plots are gated on live, single, CD45+ cells and show CD13+ CD14+ monocytes in blue, donor CD3+ T-cells in green and CD56+ CD14+ U937 target cells in orange. The percentage of respective populations in culture at baseline and after 24 h is given in the table.

Aigner, M., Feulner, J., Schaffer, S., Kischel, R., Kufer, P., Schneider, K., ... & Mackensen, A. (2013). T lymphocytes can be effectively recruited for ex vivo and in vivo lysis of AML blasts by a novel CD33/CD3-bispecific BiTE antibody construct. Leukemia, 27(5), 1107.

Figure 3 The effect of pre-stimulation of autologous T-cells on AMG330-mediated redirected lysis of primary autologous AML blasts.

AML samples were co-cultured for 144 h in the presence of 1 ng/ml AMG330 (CD33 BiTE), 1 ng/ml Control BiTE, or no BiTE. CD8+ T-cells were harvested from these cultures and incubated for the indicated time points together with a second MNC sample from the same patient at an E:T ratio of 1:1. A mean of four independent experiments from four different AML patients±s.d. is shown.

Aigner, M., Feulner, J., Schaffer, S., Kischel, R., Kufer, P., Schneider, K., ... & Mackensen, A. (2013). T lymphocytes can be effectively recruited for ex vivo and in vivo lysis of AML blasts by a novel CD33/CD3-bispecific BiTE antibody construct. Leukemia, 27(5), 1107.

Figure 1 Anti-Human CD33 Antibody in WB

Western blot analysis of TAB-756 was performed by loading Recombinant Human CD33 Protein onto a 12% Tris-HCl polyacrylamide gel. The secondary antibody: Goat Anti-Human IgG-HRP.
Lane 1: Non-Reducing Antigen (0.5 μg)
Lane 2: Reducing Antigen (0.5 μg)

Figure 2 Anti-Human CD33 Antibody in ELISA

ELISA analysis of TAB-756 was performed by coating with Recombinant Human CD33 Protein. Then blocked with BSA, and incubated with Anti-Human CD33 Antibody (TAB-756). The HRP-conjugated goat anti-human IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 3 Anti-CD33 Recombinant Antibody (Lintuzumab) (TAB-756) in SDS-PAGE

SDS-PAGE analysis of TAB-756 in non-reduced (Lane 1) and β-mercaptoethanol-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 180-245 kDa.

Figure 4 Anti-CD33 Recombinant Antibody (Lintuzumab) (TAB-756) in SEC-HPLC

HPLC analysis of TAB-756 shows that the purity is more than 95%.

Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl