Analysis of Monoclonal Antibodies Using Multiple Heart-cutting Hydrophobic Interaction/Reversed Phase 2D-LC/MS

Version 1

    Application Note

    Biotherapeutics & Biologics

     

    Abstract

    Hydrophobic interaction chromatography (HIC) is a popular approach for both downstream processing and analytical scale analysis of monoclonal antibodies (mAbs). In the latter case, HIC is a useful characterization tool because it can often separate mAb variants, for example certain isomers or oxidized species. HIC separations are also gaining popularity for the characterization of antibody-drug conjugates (ADCs). Such investigations often benefit from mass measurement using MS, but the high salt conditions used for HIC separations are completely incompatible with ESI. In this work, we overcome this obstacle using a 2D-LC system, which affords multiple heart-cutting and subsequent desalting/separation using reversed-phase chromatography on-line with TOF MS.

     

    Publication number: 5991-6376EN

    Publication Date: January 26, 2016

     

    http://www.agilent.com/cs/library/applications/5991-6376EN.pdf