Extended Data Fig. 2: Activity of enzyme candidates towards B antigens on blood group B RBCs.
Representative flow cytometry histograms of anti-B (clone 9621A8)/anti-RAMκ-Phycoerythrin (PE)-stained blood group B RBCs (filled blue) and native group O RBCs (dotted black) as staining negative controls. a Native B RBCs and b, c, d and e RBCs post-treatment with 1 μM of AmGH110A, AmGH36A, AmGH36C or AmGH27, respectively for 30 min at room temperature (RT) in conversion buffer (200 mM glycine, 3 mM NaCl, pH 6.8). f Overview of the data in a-e. The bars are means of the median fluorescence intensity (MFI) of the PE-fluorescence signal of RBC aliquots from three donors (n = 3). The open circles are the MFI values from each donor. g Native group B RBCs. h and i, RBCs from the same donor as in g, after treatment with 0.12 µM AmGH110A for 60 min at RT in conversion buffer and PBS (phosphate buffered saline), respectively. j Removal of B antigens monitored by changes of MFI at different AmGH110A concentrations over time and with RBCs from a single group B donor. k Removal of B antigens monitored by changes of MFI at different concentrations of AmGH110A in 30 min reactions with RBCs from three donors (n = 3). The MFI on y-axes in j and k are shown in logarithmic scales and the dotted line in each graph mark MFI levels of negative controls (group O RBCs). All reactions were performed with 38% haematocrit (volumetric ratio between RBCs and the source blood suspension).
