diaPASEF: parallel accumulation–serial fragmentation combined with data-independent acquisition
Data-independent acquisition modes isolate and concurrently fragment populations of different precursors by cycling through segments of a predefined precursor m/z range. Although these selection windows collectively cover the entire m/z range, overall, only a few per cent of all incoming ions are is...
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| Veröffentlicht in: | Nature methods 2020-12, Vol.17 (12), p.1229-1236 |
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| creator | Meier, Florian Brunner, Andreas-David Frank, Max Ha, Annie Bludau, Isabell Voytik, Eugenia Kaspar-Schoenefeld, Stephanie Lubeck, Markus Raether, Oliver Bache, Nicolai Aebersold, Ruedi Collins, Ben C. Röst, Hannes L. Mann, Matthias |
| description | Data-independent acquisition modes isolate and concurrently fragment populations of different precursors by cycling through segments of a predefined precursor
m/z
range. Although these selection windows collectively cover the entire
m/z
range, overall, only a few per cent of all incoming ions are isolated for mass analysis. Here, we make use of the correlation of molecular weight and ion mobility in a trapped ion mobility device (timsTOF Pro) to devise a scan mode that samples up to 100% of the peptide precursor ion current in
m/z
and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.
diaPASEF makes use of the correlation between the ion mobility and the
m
/
z
of peptides to trap and release precursor ions in a TIMS-TOF mass spectrometer for an almost complete sampling of the precursor ion beam with data-independent acquisition. |
| doi_str_mv | 10.1038/s41592-020-00998-0 |
| format | Article |
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m/z
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m/z
and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.
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m
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z
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m/z
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m/z
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m/z
and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.
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m
/
z
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m/z
range, overall, only a few per cent of all incoming ions are isolated for mass analysis. Here, we make use of the correlation of molecular weight and ion mobility in a trapped ion mobility device (timsTOF Pro) to devise a scan mode that samples up to 100% of the peptide precursor ion current in
m/z
and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.
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m
/
z
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| fulltext | fulltext |
| identifier | ISSN: 1548-7091 |
| ispartof | Nature methods, 2020-12, Vol.17 (12), p.1229-1236 |
| issn | 1548-7091 1548-7105 |
| language | eng |
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| source | MEDLINE; SpringerLink Journals; Nature |
| subjects | 631/114/2784 631/1647/296 631/45/475 Bioinformatics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biomedical Engineering/Biotechnology Cell Line, Tumor Data acquisition Data entry Data Science - methods HeLa Cells High-Throughput Screening Assays - methods Humans Ion beams Ion Channels - metabolism Ion currents Ion Transport - physiology Ionic mobility Ions Ions - chemistry Life Sciences Methods Mobility Molecular weight Peptides Precursors Proteome - metabolism Proteomes Proteomics Proteomics - methods Reproducibility of Results Tandem Mass Spectrometry - methods Time-of-flight mass spectrometry Workflow |
| title | diaPASEF: parallel accumulation–serial fragmentation combined with data-independent acquisition |
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