Development, qualification, and application of a highly efficient and robust new peak detection workflow for the...

The multi-attribute method (MAM) by liquid chromatography-mass spectrometry peptide mapping has the potential to replace multiple conventional HPLC- and capillary electrophoresis-based purity/impurity assays for release and stability testing of protein biopharmaceuticals such as monoclonal antibodies. Prerequisite is the availability of the new peak detection (NPD) functionality to reliably detect new, absent, and changed peptide species that may impair the quality, safety, and efficacy of the drug. Here, we describe the development, qualification, and application of a highly efficient and robust NPD workflow within the Genedata Expressionist® software. The detection thresholds have been rationally designed, and the NPD workflow has been successfully validated according to ICH Q2 guidelines. Individual case studies, including stability testing of drug product and detection of unknown impurities in drug substance, highlight the workflows' ability to reliably recognize relevant peptide species below 1% relative abundance without reporting any false positive peaks. The application of this NPD workflow signifies a substantial leap forward in the use of MAM as a quality control tool, as it allows identification of true positive peaks at adequate sensitivity in the absence of false positive peaks.