Human Anti-CXCR5 Recombinant Antibody (clone 40C01-VH1-Vk3) (CAT#: HPAB-0114-LSX)

Figure 1 Inhibition of CXCL13-stimulated chemotaxis of L1.2 cells expressing-human (panel A to F) and cynomolgus monkey (G and H) CXCR5, caused by anti-CXCR5 40C01-VH1-Vk3 and 40C01-VH2-Vk3 variants expressed in a whole antibody format as IgG1, IgG2 and IgG4.

The results were normalized for basal activity (medium alone) and the maximal response obtained with 10 nM CXCL13 (100%). The displayed data are representative of three independent experiments. All points were run in triplicate (error bars indicate ±S.E.M).

Figure 2 Inhibition of human CXCL13-stimulated chemotaxis of primary human B cells caused by anti-CXCR5 40C01-VH1-Vk3 (panel A) and 40C01-VH2-Vk3 (panel B) variants expressed in a whole antibody format as IgG1.

The results are expressed as calculated IC50s (half maximal inhibitory concentration). Values are the means±S.E.M from 8 (40C01-VH1-Vk3) and 5 (40C01-VH2-Vk3) separate experiments respectively.

Figure 3 Binding of Alexa-Fluor 647 labelled anti-CXCR5 40C01 variants to HEK-293 cells stably transfected with human CXCR5.

Figure 4 Binding of Alexa-Fluor 647-labelled anti-CXCR5 40C01 variants to human B cells in whole blood.

Figure 5 Binding of anti-CXCR5 40C01 variants to cynomolgus monkey B cells in whole blood.

Figure 6 Inhibition of CXCL13-stimulated ERK phosphorylation in B cells in human whole blood by anti-CXCR5 mAbs.

Figure 7 Inhibition of CXCL13-stimulated ERK phosphorylation in B cells in cynomolgus monkey whole blood by anti-CXCR5 mAbs.

Figure 8 Determination of the KD of anti-CXCR5 mAb 40C01-VH1-Vk3 variant for cell-membrane expressed CXCR5 using KinExA methodology.

CXCR5-HEK-293 cells (5×10⁶/ml) were serially diluted and incubated with 30 pM (solid diamonds) or 300 pM (open circles) active binding site concentration of anti-CXCR5 mAb 40C01-VH1-Vk3, in the presence of 0.02% NaN3 and allowed to equilibrate. The free mAb left in the supernatant was measured. (A) The % free mAb is plotted against the antigen concentration (arbitrarily taking each million cells to be equal to 10−9 M antigen). Multiple curve analysis ("n-curve analysis") using the unknown antigen method was performed to determine optimal values for KD and the antigen multiplier (B and C respectively) The 95% confidence intervals were determined by iteratively changing the optimized value for KD or antigen multiplier while keeping other parameters at their optimal values.