Best practices and benchmarks for intact protein analysis for top-down mass spectrometry

One gene can give rise to many functionally distinct proteoforms, each of which has a characteristic molecular mass. Top-down mass spectrometry enables the analysis of intact proteins and proteoforms. Here members of the Consortium for Top-Down Proteomics provide a decision tree that guides researchers to robust protocols for mass analysis of intact proteins (antibodies, membrane proteins and others) from mixtures of varying complexity. We also present cross-platform analytical benchmarks using a protein standard sample, to allow users to gauge their proficiency. The Consortium for Top-Down Proteomics presents a decision-tree-based guide to sample preparation and analysis protocols for researchers performing top-down mass-spectrometry-based analysis of intact proteins.