Characterisation of Antibody-Drug Conjugates by High-Resolution Mass Spectrometry

Antibody-drug conjugates are complex and heterogeneous molecules due to their conjugation chemistry and variable drug-to-antibody ratios (DAR). Their full characterisation requires complementary analytical techniques to assess structure, molecular weight, and conjugation patterns. High-resolution mass spectrometry provides a powerful approach to analyse ADCs at multiple levels, from intact molecules to peptides.

This application note demonstrates how HRMS enables detailed insight into ADC heterogeneity.

The full note is available for download below.

KEY HIGHLIGHTS OF THE METHOD

• Characterisation at intact, subunit, and peptide levels
• Determination of DAR and payload distribution
• Identification of positional isomers using 2D HIC-RP/UV/MS
• >99% sequence coverage for peptide mapping

VALUE FOR YOUR ADC PROGRAMME

This multi-level approach provides consistent and complementary data to better understand ADC structure and heterogeneity.

• Supports robust DAR determination across structural levels
• Enables accurate structural and molecular weight verification
• Provides insight into heterogeneity and conjugation patterns
• Facilitates identification of conjugation sites

This helps strengthen confidence in ADC characterisation.

APPLICATIONS

• Characterisation of cysteine- and lysine-conjugated ADCs
• Determination of DAR and payload distribution
• Identification of positional isomers
• Peptide mapping for conjugation site localisation