In-depth characterisation of antibody charge variants by icIEF fractionation coupled to intact mass spectrometry and peptide mapping

By
  • Thomas Menneteau
  • Claire Butré
  • Géry Van Vyncht
  • Arnaud Delobel
  • Laurence Talbot (Bio-Techne)
  • Marion Galaup (Acticor Biotech)
  • Kristell Lebozec (Acticor Biotech)
Date
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icIEF is a reference method for assessing charge variants of biotherapeutics, but it does not identify the underlying modifications. This application note presents a combined workflow integrating icIEF fractionation with intact mass spectrometry and peptide mapping to characterise these variants. Using the MauriceFlex™ system enables the collection of MS-compatible fractions for deeper analysis.

Download the full technical application note below

KEY HIGHLIGHTS OF THE METHOD

  • icIEF fractionation performed with MauriceFlex™ enabling MS-compatible fraction collection
  • Combination of icIEF, intact LC-MS and peptide mapping for variant identification
  • Fraction purity verified by re-injection into analytical icIEF
  • Identification of three modifications: pyroglutamic acid, glycation and deamidation

HOW THIS SUPPORTS YOUR ANALYTICAL STRATEGY?

This workflow enables correlation between icIEF profiles and molecular-level modifications:

  • Links charge variants to specific post-translational modifications
  • Provides MS-ready fractions without additional preparation
  • Combines intact and peptide-level analysis for better interpretation
  • Adapts to different types of modifications depending on detectability
  • Meets regulatory authority expectations as regards charge variant characterisation

TYPICAL APPLICATIONS

  • Characterisation of antibody charge variants
  • Identification of post-translational modifications
  • Characterisation of proteins showing charge heterogeneity

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