Ion-exchange chromatography, capillary isoelectric focusing, and capillary zone electrophoresis coupled to mass spectrometry for charge variant analysis of monoclonal antibodies.

ABSTRACT: Monoclonal antibody (mAb) charge variant analysis, which ensures the quality of biopharmaceuticals, is primarily performed by either cation exchange chromatography (CEX), capillary isoelectric focusing (CIEF), or capillary zone electrophoresis (CZE). Though selectivity is different for all methods, mAb proteoforms can be quantified on the intact level with a low risk of artifacts. Identification and characterization can be performed by mass spectrometry (MS), but coupling is hampered by the use of nonvolatile constituents. Here, we present comparative data and a thorough review of CEX-MS, CIEF-MS, and CZE-MS for the analysis of charge variants on the intact level. The separation of the proteoforms is confirmed by online UV detection with subsequent online MS coupling. Each method was teste

SUBMITTER: Schairer J 

PROVIDER: S-EPMC12320836 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature