2D-LC/MS Characterization of Charge Variants Using Ion Exchange and Reversed-Phase Chromatography

The determination of charge variants is one of the most important quality attributes for the characterization of biopharmaceuticals such as monoclonal antibodies (mAbs). Peak identifi cation using mass spectrometric analysis can be challenging mostly due to the incompatibility of the mass spectrometer with the buffers used during ion exchange chromatography. With the Agilent 1290 Infi nity 2D-LC solution, the second dimension enables automated desalting as well as denaturation by the addition of reversed-phase (RP) chromatography for a straightforward online LC/MS workfl ow. This Application Note demonstrates the analysis of charge variants of Rituximab (both innovator and biosimilar molecules) using multiple heart-cutting 2D-LC with MS detection.

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