T-DM1 was N-deglycosylated by incubating one unit of IgGZERO (Genovis, Lund, Sweden) per microgram of ADC for 30 min at 37 °C. For middle-up nIM-MS level experiments, IdeS digestion was performed by incubating one unit of FabRICATOR enzyme (Genovis, Lund, Sweden) per microgram of mAb or ADC for 60 min at 37 °C.

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