Ion Mobility-Mass Spectrometry and Collision Induced Unfolding Reveal Linker-Payload Effects on Antibody-Drug Conjugate Higher-Order Structure and Stability.

Key Points

• The research aims to investigate how different linker-payload configurations affect the higher-order structure and stability of antibody-drug conjugates (ADCs).
• Utilized ion mobility-mass spectrometry (IM-MS) and collision induced unfolding (CIU) techniques.
• Analyzed ADCs with various drug-to-antibody ratios (DARs) and positional isomers.
• Measured root-mean-square deviation values to evaluate higher-order structure disruption.
• Conjugation at interchain cysteine residues caused greater disruption of the ADC's higher-order structure compared to site-specific conjugates.
• Differences in conformation and stability were noted among ADCs with varying DARs.
• IM-MS and CIU proved effective as high-throughput assays for characterizing ADCs during development.