Fig. 2: Binding characteristics of DONQ52.
a Antibodies binding to Ba/F3 cells expressing various pHLA-DQ2.5 as determined by flow cytometry. Gating strategy to determine the MFI is shown in Supplementary Fig. 11a. Data are from an assay performed in triplicates (n = 3). Heat map represents relative mean MFI (%) of antibodies (DONQ52: 0.5 μg/mL, Control Ab: 10 μg/mL) to DQN0139 (10 μg/mL). Numerical data sets are shown in Supplementary Table 3. b Antibodies binding to non-HLA-DQ2.5, HLA II expressing Ba/F3 cell lines were determined by flow cytometry. Gating strategy to determine the MFI is shown in Supplementary Fig. 11a. Tu39 is anti-HLA-DR, DP, DQ antibody (mouse IgG2a). SPV-L3 is anti-pan HLA-DQ antibody (mouse IgG2a). MOPC-173 is mouse IgG2a isotype antibody. Control Ab is anti-KLH antibody (clone: IC17-SG181) as an isotype control for DONQ52 and DQN0139. Each binding antibody is colored as indicated. Data are from an assay performed in triplicates (n = 3) and are shown as mean ± SD. Numerical data sets are shown in Supplementary Table 4. c Antibodies binding to HLA-DQ2.5+ (n = 8) or HLA-DQ2.5- (n = 4) human B cells determined by flow cytometry. Relative values of MFI (%) of antibodies (DONQ52 (blue), Control Ab (gray): 10 μg/m) to DQN0139 (10 μg/mL) are shown. Peptide (-) means B cells cultured without the 33mer gliadin peptide, and Peptide (+) means B cells cultured with the 33mer gliadin peptide. Gating strategy to determine the MFI is shown in Supplementary Fig. 11b. Individual numerical data is shown in Supplementary Table 5. d Representative SPR sensorgram of the DONQ52 for binding to HLA-DQ2.5:33mer gliadin. KD value was derived by 1:1 binding model. e Representative SPR sensorgram of the DONQ52 binding to the 33mer gliadin peptide itself, or 14D5, which is an anti-gliadin antibody (Abcam PLC). f Schematic representation of DONQ52 binding properties.
