The study demonstrates fast and routine mass analysis of reduced monoclonal antibodies based on a denaturing SEC-MS method using the Waters™ BioAccord™ System.
A denaturing SEC/UV/MS method has been developed that enables the inline combination of size exclusion chromatography with optical and MS detection, for quantitative and qualitative mAb analysis at the subunit level. This well established and robust separation technique could prove useful for scientists needing a higher throughput analytical solution to meet the demand to analyze a diverse array of mAbs without customized method development. UPLC-based sub-2-μm SEC separations enabled rapid antibody light and heavy chain separations under molecule and MS-friendly conditions. This separation based on size is complementary to RP chromatography, and when coupled with MS, offers a powerful, routine and platform solution for antibody analysis.