High Resolution Antibody-Drug Conjugate Profiling with SEC and Native Mass Spectrometry

High Resolution Antibody-Drug Conjugate Profiling with SEC and Native Mass Spectrometry

Brought to you by Waters | Wyatt Technology

Overview

Antibody–drug conjugates (ADCs) present unique analytical challenges due to their structural heterogeneity and variable drug loading. One of the more unique and critical goals is accurately determining the drug‑to‑antibody ratio (DAR) because it directly influences efficacy, pharmacokinetics, and safety. Traditional DAR measurement techniques, such as UV‑based methods or hydrophobic interaction chromatography (HIC), often lack the resolution needed to fully characterize complex ADC species.

This application note shows how integrating size‑exclusion chromatography (SEC) with native mass spectrometry overcomes these limitations by enabling simultaneous assessment of glycoforms, aggregates, and drug‑loading distributions under native, label‑free conditions.

By illustrating clear drug‑distribution patterns and precise DAR values for a real‑world ADC, the app note demonstrates how SEC‑native MS provides a robust, efficient approach for comprehensive ADC characterization and faster, more reliable decision‑making in biotherapeutic development.

Key Objectives

Explore a practical case study using an FDA‑approved ADC, showcasing clear drug‑distribution patterns and precise DAR values you can use as a benchmark for your own development and QC workflows.
Learn how to confidently tackle the complexity of ADCs, including heterogeneity and precise DAR measurement, using an approach that overcomes the limitations of traditional UV and HIC methods.
See how SEC‑native MS delivers clear insight into ADC quality, enabling simultaneous assessment of glycoforms, aggregates, and drug‑loading distributions—critical for understanding efficacy and safety.
Find out how automated deconvolution and DAR reporting streamline analysis, reducing manual interpretation and improving consistency across users with different levels of LC‑MS experience.

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