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Title: High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS

Abstract

Mass spectrometric characterization of large biomolecules, such as intact proteins, requires the specificity afforded by ultra-high resolution mass measurements performed at both the intact mass and product ion levels. Although the performance of time-of-flight mass analyzers is steadily increasing, the choice of mass analyzer for large biomolecules (e.g. proteins >50 kDa) is generally limited to the Fourier transform family of mass analyzers, such as Orbitrap and ion cyclotron resonance (FTICR-MS) with the latter providing unmatched mass resolving power and measurement accuracy. Yet, protein analyses using FTMS are largely hindered by the low acquisition rates of ultra-high resolution spectra. Frequency multiple detection schemes enable FTICR-MS to overcome this fundamental barrier and achieve resolving powers and acquisition speeds 4X greater than the limits imposed by magnetic field strength. Here we expand upon earlier work on the implementation of this technique for biomolecular characterization. We report the coupling of 21T FTICR-MS, 4X frequency multiplication, ion trapping field harmonization technology, and spectral data processing methods to achieve unprecedented acquisition rates and resolving power in mass spectrometry of large intact proteins. Isotopically resolved spectra of multiply charged ubiquitin ions were acquired using detection periods as short as 12 ms. Large proteins such as apo-transferrin (MW=78more » kDa) and monoclonal antibody (MW=150 kDa) were isotopically resolved with detection periods of 384 ms and 768 ms, respectively. These results illustrate the future capability of accurate characterization of large proteins on time-scales compatible with online separations.« less

Authors:
 [1];  [2]; ORCiD logo [1];  [1]
  1. BATTELLE (PACIFIC NW LAB)
  2. Russian Academy of Sciences
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1557725
Report Number(s):
PNNL-SA-133517
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry
Additional Journal Information:
Journal Volume: 90; Journal Issue: 9
Country of Publication:
United States
Language:
English

Citation Formats

Shaw, Jared B., Gorshkov, Mikhail, Wu, Qinghao, and Pasa Tolic, Ljiljana. High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS. United States: N. p., 2018. Web. doi:10.1021/acs.analchem.7b04606.
Shaw, Jared B., Gorshkov, Mikhail, Wu, Qinghao, & Pasa Tolic, Ljiljana. High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS. United States. doi:10.1021/acs.analchem.7b04606.
Shaw, Jared B., Gorshkov, Mikhail, Wu, Qinghao, and Pasa Tolic, Ljiljana. Tue . "High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS". United States. doi:10.1021/acs.analchem.7b04606.
@article{osti_1557725,
title = {High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS},
author = {Shaw, Jared B. and Gorshkov, Mikhail and Wu, Qinghao and Pasa Tolic, Ljiljana},
abstractNote = {Mass spectrometric characterization of large biomolecules, such as intact proteins, requires the specificity afforded by ultra-high resolution mass measurements performed at both the intact mass and product ion levels. Although the performance of time-of-flight mass analyzers is steadily increasing, the choice of mass analyzer for large biomolecules (e.g. proteins >50 kDa) is generally limited to the Fourier transform family of mass analyzers, such as Orbitrap and ion cyclotron resonance (FTICR-MS) with the latter providing unmatched mass resolving power and measurement accuracy. Yet, protein analyses using FTMS are largely hindered by the low acquisition rates of ultra-high resolution spectra. Frequency multiple detection schemes enable FTICR-MS to overcome this fundamental barrier and achieve resolving powers and acquisition speeds 4X greater than the limits imposed by magnetic field strength. Here we expand upon earlier work on the implementation of this technique for biomolecular characterization. We report the coupling of 21T FTICR-MS, 4X frequency multiplication, ion trapping field harmonization technology, and spectral data processing methods to achieve unprecedented acquisition rates and resolving power in mass spectrometry of large intact proteins. Isotopically resolved spectra of multiply charged ubiquitin ions were acquired using detection periods as short as 12 ms. Large proteins such as apo-transferrin (MW=78 kDa) and monoclonal antibody (MW=150 kDa) were isotopically resolved with detection periods of 384 ms and 768 ms, respectively. These results illustrate the future capability of accurate characterization of large proteins on time-scales compatible with online separations.},
doi = {10.1021/acs.analchem.7b04606},
journal = {Analytical Chemistry},
number = 9,
volume = 90,
place = {United States},
year = {2018},
month = {5}
}