A standardised LC-MS workflow for rapid DAR determination of thiol-conjugated ADCs across a variety of toxin linkers

Fast, reliable and generic methods are required for drug to antibody ratio (DAR) determination in an ADC development laboratory. In an effort to establish such a method, we have expanded on a previously developed “middle-up” workflow for determination of DAR by liquid chromatography- mass spectrometry (LC-MS). We demonstrate application of the workflow to a range of stochastically conjugated ADCs with DARs from 1 – 8, incorporating auristatin, α-amanitin, duocarmycin, PBD dimer and DX8951 payloads.

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