Abstract

Top-down proteomics is the analysis of intact proteins in their endogenous form without proteolysis, preserving valuable information about post-translation modifications, isoforms and proteolytic processing. The quality of top-down LC-MS/MS datasets is rapidly increasing due to advances in instrumentation and sample processing protocols. However, the top-down mass spectra are substantially more complex compared to conventional bottom-up data. To take full advantage of the increasing data quality, there is an urgent need to develop algorithms and software tools for confident proteoform identification and quantification. In this study, we present a new open source software suite for top-down proteomics analysis consisting of an LC-MS feature finding algorithm, a database search algorithm, and an interactive results viewer. The presented tool along with several other popular tools were evaluated using human-in-mouse xenograft luminal and basal breast tumor samples that are known to have significant differences in protein abundance based on bottom-up analysis.

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