Narrow-window DIA: Ultra-fast quantitative analysis of comprehensive proteomes with high sequencing depth
Abstract Mass spectrometry (MS)-based proteomics aims to characterize comprehensive proteomes in a fast and reproducible manner. Here, we present an ultra-fast scanning data-independent acquisition (DIA) strategy consisting on 2-Th precursor isolation windows, dissolving the differences between data-dependent and independent methods. This is achieved by pairing a Quadrupole Orbitrap mass spectrometer with the asymmetric track lossless (Astral) analyzer that provides >200 Hz MS/MS scanning speed, high resolving power and sensitivity, as well as low ppm-mass accuracy. Narrow-window DIA enables profiling of up to 100 full yeast proteomes per day, or ∼10,000 human proteins in half-an-hour. Moreover, multi-shot acquisition of fractionated samples allows comprehensive coverage of human proteomes in ∼3h, showing comparable depth to next-generation RNA sequencing and with 10x higher throughput compared to current state-of-the-art MS. High quantitative precision and accuracy is demonstrated with high peptide coverage in a 3-species proteome mixture, quantifying 14,000+ proteins in a single run in half-an-hour.Teaser Accurate and precise label-free quantification with comprehensive proteome coverage using narrow-window DIA.
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Preprint| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 10. Juni Zur Gesamtaufnahme - year:2023 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Guzman, Ulises H [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2023.06.02.543374 |
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| 520 | |a Abstract Mass spectrometry (MS)-based proteomics aims to characterize comprehensive proteomes in a fast and reproducible manner. Here, we present an ultra-fast scanning data-independent acquisition (DIA) strategy consisting on 2-Th precursor isolation windows, dissolving the differences between data-dependent and independent methods. This is achieved by pairing a Quadrupole Orbitrap mass spectrometer with the asymmetric track lossless (Astral) analyzer that provides >200 Hz MS/MS scanning speed, high resolving power and sensitivity, as well as low ppm-mass accuracy. Narrow-window DIA enables profiling of up to 100 full yeast proteomes per day, or ∼10,000 human proteins in half-an-hour. Moreover, multi-shot acquisition of fractionated samples allows comprehensive coverage of human proteomes in ∼3h, showing comparable depth to next-generation RNA sequencing and with 10x higher throughput compared to current state-of-the-art MS. High quantitative precision and accuracy is demonstrated with high peptide coverage in a 3-species proteome mixture, quantifying 14,000+ proteins in a single run in half-an-hour.Teaser Accurate and precise label-free quantification with comprehensive proteome coverage using narrow-window DIA | ||
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| 700 | 1 | |a Martinez Del Val, Ana |0 (orcid)0000-0002-8784-3822 |4 aut | |
| 700 | 1 | |a Ye, Zilu |0 (orcid)0000-0001-8829-6579 |4 aut | |
| 700 | 1 | |a Damoc, Eugen |4 aut | |
| 700 | 1 | |a Arrey, Tabiwang N. |4 aut | |
| 700 | 1 | |a Pashkova, Anna |4 aut | |
| 700 | 1 | |a Denisov, Eduard |4 aut | |
| 700 | 1 | |a Petzoldt, Johannes |4 aut | |
| 700 | 1 | |a Peterson, Amelia C. |4 aut | |
| 700 | 1 | |a Harking, Florian |0 (orcid)0000-0001-5553-1421 |4 aut | |
| 700 | 1 | |a Østergaard, Ole |4 aut | |
| 700 | 1 | |a Stewart, Hamish |4 aut | |
| 700 | 1 | |a Xuan, Yue |4 aut | |
| 700 | 1 | |a Hermanson, Daniel |4 aut | |
| 700 | 1 | |a Hock, Christian |4 aut | |
| 700 | 1 | |a Makarov, Alexander |0 (orcid)0000-0002-7046-6709 |4 aut | |
| 700 | 1 | |a Zabrouskov, Vlad |4 aut | |
| 700 | 1 | |a Olsen, Jesper V. |0 (orcid)0000-0002-4747-4938 |4 aut | |
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