Labile post-translational modifications are better preserved using ECD than conventional MS fragmentation techniques such as collision-induced dissociation (CID). Common modifications, such as glycosylation and phosphorylation, can impact the safety, efficacy, and binding activity of monoclonal antibodies but are challenging to identify. Stephen Sciuto from Agilent Technologies and colleagues analyzed tryptic digests of NIST mAb and Infliximab using an 6545XT Q-TOF mass spectrometer in ExD or CID mode. ECD provided higher sequence coverage than CID, reaching 100% for glycosylated peptides in Infliximab. The ability to pinpoint glycosylation sites using fragment ions demonstrates the power of ECD for characterizing glycopeptides.
Analyze Charge Variants in Antibody-Based Therapeutics Using Cation Exchange Chromatography
July 18th 2024This application note discusses the latest development on high performance cation exchange bio-separation columns for determining charge variants in protein-based biotherapeutics, as well as the strategy on method development.
The Power of HPLC/SFC Control and AQbD in One Software
July 17th 2024Download this infographic for a quick overview of Shimadzu’s LabSolutions Method Development (MD) software, an effective and automated software solution for U/HPLC method development that uses analytical quality by design (AQbD) principles in its workflow.