Figure 1: The binding properties of HS20 monoclonal antibody.
From: Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
(a) The schematic structure of GPC3, the human monoclonal antibody HS20 recognizing the heparan sulfate chains on GPC3 and the mouse monoclonal antibody YP7 recognizing a C-terminal region of the GPC3 core protein. (b) HS20 did not bind the mutant GPC3 without heparan sulfate chains (GPC3ΔHS). ELISA results showed the binding properties of HS20 and YP7 on wild type GPC3 and GPC3ΔHS. BSA was used as the negative antigen control. Values represent mean ± SD, t-test. (c) The binding property of HS20 to various glypicans. ELISA results show the binding affinities of HS20 for GPC1, GPC3, GPC5 and GPC6. The X-axis represents the concentration of HS20, while the Y axis indicates protein binding as determined by the OD450 nm. (d) HS20 lost binding capacity on the cells without heparan sulfate. Flow results showed the binding of HS20 for CHO-K1 cells and pgsA-745 cells, which lack the ability to produce HS and CS. (e) HS but not CS inhibited HS20 binding for GPC3. A competitive ELISA where HS20 (5 μg/ml) was pre-incubated with increasing concentrations of HS or bikunin (CS) for 30 minutes, and then added to an ELISA plate to detect the binding to GPC3.
