2D-LC/MS Characterization of Charge Variants Using Ion Exchange and Reversed-Phase Chromatography Multiple Heart-Cutting 2D-LC Analysis of Innovator versus Biosimilar Monoclonal Antibodies

Version 2

    Application Note

    Biotherapeutics and Biologics



    The determination of charge variants is one of the most important quality attributes for the characterization of bio pharmaceuticals such as monoclonal antibodies (mAbs). Peak identification 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 Infinity 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 workflow. 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.


    Publication number: 5991-6673EN

    Publication Date: May 1, 2016