Decoding IgG2 Disulfide Isoforms with Native CEX-MS and Advanced Peptide Mapping

About this webinar

Join Yann Leblanc from Servier R&D as he shares insights into characterizing IgG2 disulfide isoforms using native cation exchange chromatography-mass spectrometry (CEX-MS) combined with advanced peptide mapping. The IgG2 subclass presents unique structural complexity—like a molecular blueprint with multiple architectural variants—where disulfide-bonded isoforms (A, A/B, and B) form different structural scaffolds that may influence biological function and therapeutic activity.

This presentation demonstrates the analytical framework for mapping these structural variations. Using native LC-MS workflows integrated with Protein Metrics' Byos and Byonic software, the approach enables high-resolution separation and characterization of IgG2 disulfide isoforms while preserving their native structure. Discover how this method resolves, identifies, and quantifies these critical structural variants directly under near-physiological conditions, providing the foundation for therapeutic antibody development.

Key Takeaways:

1. Native CEX-MS Workflow: See how volatile salts and pH-gradient Cation Exchange Chromatography preserve native antibody structure while separating F(ab′)₂ fragments for high-quality mass spectra.
2. Automated Data Analysis: Learn how Byos and Byonic software streamline intact mass deconvolution and disulfide bond assignment using the Isotope Envelope Confidence (IEC) scoring system.
3. Complete Structural Mapping: Discover how middle-up strategies with peptide mapping provide detailed profiling of disulfide bonds, revealing molecular heterogeneity important for quality control and biosimilar comparability.

Why This Matters

Disulfide bond heterogeneity in IgG2 is a critical quality attribute influencing antibody biological functions. Having the right analytical blueprint enables biopharmaceutical scientists to monitor and control product quality during manufacturing, understand the structural framework of isoform formation and rearrangement, support biosimilar comparability studies, and build more effective therapeutic antibody development pipelines.