FLASHIda enables intelligent data acquisition for top–down proteomics to boost proteoform identification counts

Kyowon Jeong (), Maša Babović, Vladimir Gorshkov, Jihyung Kim, Ole N. Jensen and Oliver Kohlbacher ()
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Kyowon Jeong: University of Tübingen
Maša Babović: University of Southern Denmark
Vladimir Gorshkov: University of Southern Denmark
Jihyung Kim: University of Tübingen
Ole N. Jensen: University of Southern Denmark
Oliver Kohlbacher: University of Tübingen

Nature Communications, 2022, vol. 13, issue 1, 1-12

Abstract: Abstract The detailed analysis and structural characterization of proteoforms by top-down proteomics (TDP) has gained a lot of interest in biomedical research. Data-dependent acquisition (DDA) of intact proteins is non-trivial due to the diversity and complexity of proteoforms. Dedicated acquisition methods thus have the potential to greatly improve TDP. Here, we present FLASHIda, an intelligent online data acquisition algorithm for TDP that ensures the real-time selection of high-quality precursors of diverse proteoforms. FLASHIda combines fast charge deconvolution algorithms and machine learning-based quality assessment for optimal precursor selection. In an analysis of E. coli lysate, FLASHIda increases the number of unique proteoform level identifications from 800 to 1500 or generates a near-identical number of identifications in one third of the instrument time when compared to standard DDA mode. Furthermore, FLASHIda enables sensitive mapping of post-translational modifications and detection of chemical adducts. As a software extension module to the instrument, FLASHIda can be readily adopted for TDP studies of complex samples to enhance proteoform identification rates.

Date: 2022
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DOI: 10.1038/s41467-022-31922-z

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