Improvements on sample preparation and peptide separation for reduced peptide mapping based multi-attribute method analysis of therapeutic monoclonal antibodies using lysyl endopeptidase digestion

•The first high-resolution MS based Multi-Attribute Method (MAM) platform using Lys-C digestion without desalting.•The first MAM platform with improved chromatography separation of hydrophilic short peptides, including the stability indicating VSNK peptide.•Automated sample preparation using Lys-C digestion without desalting had similar performance as manual digestion for potential high-throughput MAM testing.•Demonstrated robustness of the MAM workflow with Lys-C digestion without desalting.