Figure 5.

Qualification of the proposed method using 0.15% DFA-modified mobile phases with different cysteine-linked ADCs. (a) Subunits from a cysteine-linked auristatin conjugated antibody previously used in the proposed method. (b) Subunits from a commercially available cysteine linked, dansyl-cadaverine-SMCC conjugated ADC mimic. (c) Subunits from a second cysteine-linked ADC, separated using higher temperature. Separations were performed with a 2.1 × 150 mm column packed with a high coverage phenyl bonded 2.7 µm superficially porous 450 Å stationary phase using a flow rate of 0.6 mL/min, column temperature of 70°C or 80°C, and 1 μg mass loads.