Human Anti-FCGR2B Recombinant Antibody (clone 6G08) (CAT#: PABS-0156)

Figure 1 Binding profile of mAbs on human leukocyte populations.

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 2 Dose-dependent binding of mAbs to B cells.

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 3 Ability of N297Q hFcgRIIB mAbs to elicit (A) or block (B) hFcgRIIB ITIM phosphorylation.

(A) Ability of N297Q hFcgRIIB mAbs to elicit hFcgRIIB ITIM phosphorylation (pFcgRIIB) on Raji cells; hIgG1 isotype control (iso ctrl) and Rit used as negative and positive controls, respectively. a-Tubulin loading control.
(B) Ability of N297Q hFcgRIIB mAbs to block hFcgRIIB ITIM phosphorylation induced by Rit. Representative blots (n = 3) shown in (A) and (B).

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 4 Ability of N297Q hFcgRIIB mAbs to block internalization induced by Rit.

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 5 Correlation between the ability of hFcgRIIB mAbs to block Rit internalization (shown in C) and their relative ranked affinities (R2 = 0.78).

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 6 Correlation between the ability to block Rit internalization and the ability to block Rit-induced phosphorylation of hFcgRIIB (shown in B) (R2 = 0.79).

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 7 hFcγRIIB-specific mAbs were incubated with human peripheral blood leukocytes and then assessed for binding to CD14⁺ monocytes (D), CD19⁺ B cells (E) or CD3⁺ T cells (F) using a secondary mAb by flow cytometry and percentage positive cells plotted (± SEM of 3 healthy blood donors).

Clone 6A09 showed cross-reactivity towards both monocytes and B cells (D and E)

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 8 Ability of WT and/or N297Q (NQ) hFcγRIIB specific mAbs (clones 6G11 and 6G08) to elicit hFcγRIIB ITIM phosphorylation (pFcγRIIB)

(A-B) Ability of WT and/or N297Q (NQ) hFcγRIIB specific mAbs (clones 6G11 and 6G08) to elicit hFcγRIIB ITIM phosphorylation (pFcγRIIB) on isolated human tonsil B cells (A) and primary peripheral blood monocytes (B), respectively. α-Tubulin, GAPDH and hFcγRIIB were used as loading controls, as indicated; representative blots shown.

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 9 hFcγRIIBtransfected Ramos cells were treated with WT or N297Q variants (10 µg/ml) of a selection of hFcγRIIB mAbs

hFcγRIIBtransfected Ramos cells were treated with WT or N297Q variants (10 µg/ml) of a selection of hFcγRIIB mAbs (representing a selection of antagonists and agonists) or isotype control (iso ctrl) and AF488-labeled Rit (5 µg/ml) for the indicated time-points at 37°C. Internalization of Rit was determined using a quenching assay and expressed as the percentage of surface CD20 expression. AF488-labeled tositumomab (type II CD20 mAb) was used as negative control, as it does not rapidly internalize following engagement of CD20 (Beers et al., 2010).

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.

Figure 10 ADDC activity

Raji cells pre-opsonized with hFcγRIIB mAbs were co-cultured with NK cells purified from the peripheral blood of healthy donors and ADDC activity was assessed.

Roghanian, A., Teige, I., Mårtensson, L., Cox, K. L., Kovacek, M., Ljungars, A., ... & Cragg, M. S. (2015). Antagonistic human FcγRIIB (CD32B) antibodies have anti-tumor activity and overcome resistance to antibody therapy in vivo. Cancer cell, 27(4), 473-488.