Abstract

The ratio of the conjugated drug to mAb (drug-to-antibody ratio or DAR) is one of the critical quality attributes for ADC development because it can affect efficacy and safety. This talk highlight a very simple workflow solution for a very complex challenge of error prone sample preparation such as affinity capture of ADCs from complex matrices (for example, serum/plasma) and deglycosylation of the purified antibodies. The accurate measurement of these DAR values were calculated using data generated by a Hi Resolution Mass Spectrometery. The whole automated process, including antigen immobilization, affinity purification, and on-cartridge deglycosylation, can be completed in approximately five hours, with minimal hands-on time. The techniques involved in the workflow are AssayMAP Bravo with an Agilent 6545XT AdvanceBio LC/Q-TOF, and Agilent BioConfirm DAR Calculator software.