Abstract
Top-down proteomics is a key mass spectrometry-based technology for comprehensive analysis of proteoforms. Proteoforms exhibit multiple high charge states and isotopic forms in full MS scans. The dissociation behavior of proteoforms in different charge states and subjected to different collision energies is highly variable. The current widely employed data-dependent acquisition (DDA) method selects a narrow m/z range (corresponding to a single proteoform charge state) for dissociation from the most abundant precursors. We describe here Mesh, a novel dissociation strategy, to dissociate multiple charge states of one proteoform with multiple collision energies. We show that the Mesh strategy has the potential to generate fragment ions with improved sequence coverage and improve identification ratios in top-down proteomic analyses of complex samples. The strategy is implemented within an open-source instrument control software program named MetaDrive to perform real time deconvolution and precursor selection.
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