Here we utilize a cysteine-linked antibody fluorophore conjugate to show how LC-MS methods may be optimized using a non-toxic ADC-mimic. The ADC-mimic was used to develop native and reduced SEC-MS conditions. The reduced mimic’s heavy and light chains, as well as its IdeS-proteolysis fragments, were used to develop reversed-phase LC-MS methods. A tryptic digest of the ADC mimic was used to develop a peptide mapping LC-MS method.
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