Measurement of Individual Ions Sharply Increases the Resolution of Orbitrap Mass Spectra of Proteins

Jared O. Kafader, Rafael D. Melani, Michael W. Senko, Alexander A. Makarov, Neil L Kelleher, Philip D Compton*

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

It is well-known that with Orbitrap-based Fourier-transform-mass-spectrometry (FT-MS) analysis, longer-time-domain signals are needed to better resolve species of interest. Unfortunately, increasing the signal-acquisition period comes at the expense of increasing ion decay, which lowers signal-to-noise ratios and ultimately limits resolution. This is especially problematic for intact proteins, including antibodies, which demonstrate rapid decay because of their larger collisional cross-sections, and result in more frequent collisions with background gas molecules. Provided here is a method that utilizes numerous low-ion-count spectra and single-ion processing to reconstruct a conventional m/z spectrum. This technique has been applied to proteins varying in molecular weight from 8 to 150 kDa, with a resolving power of 677»000 achieved for transients of carbonic anhydrase (29 kDa) with a duration of only 250 ms. A resolution improvement ranging from 10-to 20-fold was observed for all proteins, providing isotopic resolution where none was previously present.

Original languageEnglish (US)
Pages (from-to)2776-2783
Number of pages8
JournalAnalytical Chemistry
Volume91
Issue number4
DOIs
StatePublished - Feb 19 2019

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Ions
Proteins
Carbonic Anhydrases
Optical resolving power
Mass spectrometry
Signal to noise ratio
Fourier transforms
Gases
Molecular weight
Molecules
Antibodies
Processing

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Kafader, Jared O. ; Melani, Rafael D. ; Senko, Michael W. ; Makarov, Alexander A. ; Kelleher, Neil L ; Compton, Philip D. / Measurement of Individual Ions Sharply Increases the Resolution of Orbitrap Mass Spectra of Proteins. In: Analytical Chemistry. 2019 ; Vol. 91, No. 4. pp. 2776-2783.
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Measurement of Individual Ions Sharply Increases the Resolution of Orbitrap Mass Spectra of Proteins. / Kafader, Jared O.; Melani, Rafael D.; Senko, Michael W.; Makarov, Alexander A.; Kelleher, Neil L; Compton, Philip D.

In: Analytical Chemistry, Vol. 91, No. 4, 19.02.2019, p. 2776-2783.

Research output: Contribution to journalArticle

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