Article: LC-UV-MS method development for antibody drug conjugates using a non-toxic ADC-mimic

Antibody-drug conjugates (ADC) represent a rapidly emerging class of biotherapeutic molecules, which harness the specificity of monoclonal antibodies (mAb) to selectively deliver highly potent cytotoxic drugs. Like all biotherapeutics they require significant analytical characterization.

A non-toxic ADC surrogate, or “ADC Mimic,” with highly similar physicochemical properties to commercial ADCs provides an excellent molecule to safely drive analytical method development for ADCs.

ADC Mimic – MSQC4 plus 2, 4, 6, or 8 dansylcadaverine- SMCC mimic-linkers. Each conjugation to a Cys residue imparts a +668 Da mass shift.
Here, we build on previous work describing the preparation of such a non-toxic ADC-Mimic by utilizing the ADC-Mimic and the underlying mAb (MSQC4) to perform a comparison of chromatographic stationary phases. We also present the use of the ADCMimic to develop a workflow to characterize ADC variants by first separating them via hydrophobic interaction chromatography (HIC), collecting fractions, and subsequently analyzing the ADC variants by RP-LC-UV-MS.

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