English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Online parallel accumulation - serial fragmentation (PASEF) with a novel trapped on mobility mass spectrometer

Meier, F., Brunner, A.-D., Koch, S., Koch, H., Lubeck, M., Krause, M., et al. (2018). Online parallel accumulation - serial fragmentation (PASEF) with a novel trapped on mobility mass spectrometer. Molecular and Cellular Proteomics, 17(12): UNSP TIR118.000900, pp. 2533-2545. doi:10.1074/mcp.TIR118.000900.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/21.11116/0000-0002-DE9E-3 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-CA8F-9
Genre: Journal Article

Files

show Files
hide Files
:
2534.full.pdf (Publisher version), 4MB
Name:
2534.full.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
© 2018 Meier et al.

Locators

show
hide
Locator:
https://www.ebi.ac.uk/pride/archive/ (Supplementary material)
Description:
The raw datasets and the MaxQuant output files generated and analyzed throughout this study have been deposited at the ProteomeXchange Consortium via the PRIDE partner repository (55) with the dataset identifier PXD010012 (https://www.ebi.ac.uk/pride/archive/). MaxQuant msms.txt and .apl files are provided for viewing of annotated spectra, including spectra of single peptide protein identifications.

Creators

show
hide
 Creators:
Meier, Florian1, Author              
Brunner, Andreas-David1, Author              
Koch, Scarlet2, Author
Koch, Heiner2, Author
Lubeck, Markus2, Author
Krause, Michael2, Author
Goedecke, Niels2, Author
Decker, Jens2, Author
Kosinski, Thomas2, Author
Park, Melvin A.2, Author
Bache, Nicolai2, Author
Hoerning, Ole2, Author
Cox, Jürgen3, Author              
Raether, Oliver2, Author
Mann, Matthias1, Author              
Affiliations:
1Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              
2external, ou_persistent22              
3Cox, Jürgen / Computational Systems Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_2063284              

Content

show
hide
Free keywords: ION; FUNDAMENTALS; PROTEOMICS; MIXTURES; STRATEGY; PLATFORM; DATABASE; RANGEBiochemistry & Molecular Biology;
 Abstract: In bottom-up proteomics, peptides are separated by liquid chromatography with elution peak widths in the range of seconds, whereas mass spectra are acquired in about 100 microseconds with time-of-flight (TOF) instruments. This allows adding ion mobility as a third dimension of separation. Among several formats, trapped ion mobility spectrometry (TIMS) is attractive because of its small size, low voltage requirements and high efficiency of ion utilization. We have recently demonstrated a scan mode termed parallel accumulation - serial fragmentation (PASEF), which multiplies the sequencing speed without any loss in sensitivity (Meier et aL, PMID: 26538118). Here we introduce the timsTOF Pro instrument, which optimally implements online PASEF. It features an orthogonal ion path into the ion mobility device, limiting the amount of debris entering the instrument and making it very robust in daily operation. We investigate different precursor selection schemes for shotgun proteomics to optimally allocate in excess of 100 fragmentation events per second. More than 600,000 fragmentation spectra in standard 120 min LC runs are achievable, which can be used for near exhaustive precursor selection in complex mixtures or accumulating the signal of weak precursors. In 120 min single runs of HeLa digest, Max-Quant identified more than 6,000 proteins without matching to a library and with high quantitative reproducibility (R > 0.97). Online PASEF achieves a remarkable sensitivity with more than 2,500 proteins identified in 30 min runs of only 10 ng HeLa digest. We also show that highly reproducible collisional cross sections can be acquired on a large scale (R > 0.99). PASEF on the timsTOF Pro is a valuable addition to the technological toolbox in proteomics, with a number of unique operating modes that are only beginning to be explored.

Details

show
hide
Language(s): eng - English
 Dates: 2018-12-14
 Publication Status: Published online
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000452476000018
DOI: 10.1074/mcp.TIR118.000900
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Molecular and Cellular Proteomics
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Bethesda, MD : American Society for Biochemistry and Molecular Biology
Pages: - Volume / Issue: 17 (12) Sequence Number: UNSP TIR118.000900 Start / End Page: 2533 - 2545 Identifier: ISSN: 1535-9476
CoNE: https://pure.mpg.de/cone/journals/resource/111035577487002