Fig. 8
B12–drug conjugate enhances the cytotoxic effect of the native antibody. a Antibody in its native form was reduced using 30 equivalents DTT for 3 h at 37 °C in PBS, to reduce the interchain disulfide bonds. The reduced cysteines were then reacted with 60 equivalents of carbonyl acrylic acid–valine-citrulline-monomethyl auristatin E (compound A) for up to 24 h at 37 °C. The average Drug-to-Antibody ratio (DAR) was 4. b HPB-ALL cells, c D1mutP2 and control D1 parental cells, d primary T-ALL cells collected from diagnostic patients, and e PDX samples were treated with B12 or B12–drug conjugate at the indicated concentrations. Cell death was analyzed after 72 h of culture, using Annexin V/7AAD staining and data was analyzed by flow cytometry. Percentage of live cells was calculated by normalizing to the untreated cells
