Fig. 1

Analysis of hOA-DN30 structure and function. a Schematic representation of hOA-DN30. VH: humanized variable domain of the heavy chain. CH1, CH2, CH3: constant domains derived from human IgG1 heavy chain. VL: humanized variable domain of the light chain. CL: constant domain derived from human Igk light chain. b SDS-PAGE in polyacrylamide gel under non-reducing (NR) and reducing (R) conditions, followed by staining with GelCode Blue Stain reagent. MW: Molecular Weight; Hc: Heavy chain; Lc Light chain; Fc: fragment crystallizable region. c ELISA binding analysis of hOA-DN30 and MvDN30 (liquid phase) to a Met-Fc chimera (solid phase). OD, optical density at 450 nm. Dissociation constant values (Kd) ± SD are reported in the graph. Each point is the mean of triplicate values; bars represent SD. d HPAF-II cells untreated (Ctrl) or treated with HGF (8 ng/ml), or bivalent murine DN30 mAb (200 nM), or hOA-DN30 (200 nM) for 18 h. Pictures are representative of the cell phenotype: control cells grew in compact islands, while HGF-treated cells appeared as single cells separated from each other (scattered). Some scattered islands were found in the chimeric DN30 mAb treated cells. Stimulation with hOA-DN30 did not induce cell motility, appearing indistinguishable from controls. Data reported in the figure are representative of at least two experiments done