This technical note demonstrates a novel integrated workflow using the Intabio ZT system for charge variant analysis of antibody-drug conjugates (ADCs). This innovative system offers direct chip-based integration of icIEF with mass spectrometry (MS), achieving separation and confident identification of proteoforms with different payloads, reliable quantitation of charge variants and determination of drug-to-antibody ratios (DARs). Further, the streamlined icIEF-UV/MS platform significantly reduces acquisition and analysis time compared to ion exchange chromatography (IEX) with fraction collection, eliminating multiple weeks of effort.1
The icIEF-UV/MS data were interpreted using an Intabio intact icIEF-UV/MS workflow within Biologics Explorer software for automated protein deconvolution and accurate DAR calculation. With a single injection, 8 different trastuzumab emtansine (T-DM1) payloads were electrophoretically separated and an average DAR of 3.6 was measured.
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