Charge heterogeneity characterisation of an IgG4-based mAb using AEX coupled to MS

Cation exchange chromatography (CEX) is very well suited for characterising the charge heterogeneity of biomolecules. This also applies to most commercially available monoclonal antibodies (mAbs). They are often based on IgG1 and have a high isoelectric point (pI). Anion exchange chromatography (AEX), has so far only been used for relatively acidic proteins.

In addition, IgG4-based mAbs are becoming increasingly important. Four FDA-approved mAbs are already based on IgG4 and at least six other candidates are in development [1]. This application demonstrates that the charge heterogeneity of an IgG4-based mAb with a pI of 6.6 (Regeneron’s proprietary mAb) can be successfully evaluated using a strong anion exchange column (SAX), BioPro IEX QF, in conjunction with MS detection [2].

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