Anti-Human BAFF Recombinant Antibody (Tabalumab) (CAT#: TAB-216)

Figure 1 Tabalumab binds membrane-bound BAFF on transfected cells or activated human monocytes.

(A) HEK293 cells were stably transfected with non-cleavable human BAFF. Membrane-bound BAFF was detected with Alexa 488-labeled tabalumab (gray histogram) compared with Alexa 488-labeled isotype control (black histogram). (B) Purified human CD14+ monocytes were activated with IFN-γ for 3 days. Membrane-bound BAFF was detected with Alexa 488-labeled tabalumab (gray histogram) compared with Alexa 488-labeled isotype control (black histogram). Data are from one experiment but representative of n=12 experiments (A) or ten donors (B).

Manetta, J., Bina, H., Ryan, P., Fox, N., Witcher, D. R., & Kikly, K. (2014). Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor. Journal of inflammation research, 7, 121.

Figure 2 Tabalumab neutralization of soluble BAFF from multiple species.

IC50 was calculated for soluble human BAFF (IC50: 104 pM, 95% CI 96–112 pM), soluble cynomolgus monkey BAFF (IC50: 143 pM, 95% CI 126–162 pM), or soluble rabbit BAFF (IC50: 176 pM, 95% CI 170–182 pM). T1165.17 cells were stimulated with 365 pM BAFF and proliferation was measured using a colorimetric method for determining the number of viable cells at 44 hours. Data are presented as mean ± SD.

Manetta, J., Bina, H., Ryan, P., Fox, N., Witcher, D. R., & Kikly, K. (2014). Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor. Journal of inflammation research, 7, 121.

Figure 3 Administration of tabalumab to human BAFF transgenic mice leads to decreased B-cells and a reduction in non-canonical NF-κB signaling.

Tabalumab (■) or isotype control (○) was injected SC on day 0. Mice were sacrificed on days 2, 4, 8, 11, 16, and 23. (A) Splenic B-cells were enumerated by flow cytometry using B220; (B) p52 levels in the spleen were evaluated by a binding assay; (C) Western blot showing bands for p100 and p52 in splenocyte lysates; Grb2 was monitored to ensure equivalent loading of wells. * represents P-value <0.05. (●) represents an untreated non-transgenic mouse.

Manetta, J., Bina, H., Ryan, P., Fox, N., Witcher, D. R., & Kikly, K. (2014). Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor. Journal of inflammation research, 7, 121.

Figure 1 Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in SDS-PAGE

Lane 1: Reduced mAb
Lane 2: Non-reduced mAb

Figure 2 Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in SEC-HPLC

The purity of TAB-216-ADCP was greater than 95% as determined by SEC-HPLC.

Figure 3 Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in WB

Western blot analysis of TAB-216-ADCP was performed by loading recombinant Human BAFF with his-tag.

TAB-216-ADCP incubation concentration: 2 ng/μL.
The secondary antibody: HRP-Goat anti-human IgG
Lane 1: Reduced Antigen (0.1 μg)
Lane 2: Reduced Antigen (0.3 μg)
Lane 3: Reduced Antigen (0.6 μg)

Figure 4 Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in ELISA

ELISA analysis of TAB-216-ADCP was performed by coating with recombinant Human BAFF with his-tag.

The secondary antibody: HRP-Goat anti-human IgG

Figure 5 Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in DB

Anti-Human BAFF Recombinant Antibody (Tabalumab), ADCP enhanced (TAB-216-ADCP) in DB

Dot Blot analysis of TAB-216-ADCP was performed by coating with recombinant Human BAFF with his-tag.

The secondary antibody: HRP-Goat anti-human IgG