Integration of Segmented Ion Fractionation and Differential Ion Mobility on a Q-Exactive Hybrid Quadrupole Orbitrap Mass Spectrometer.

Autor:	Pfammatter S; Institute for Research in Immunology and Cancer (IRIC)Université de Montréal, Montréal, Quebec H3C 3J7, Canada.; Department of Chemistry, Université de Montréal, Montréal, Quebec H3C 3J7, Canada., Wu Z; Institute for Research in Immunology and Cancer (IRIC)Université de Montréal, Montréal, Quebec H3C 3J7, Canada.; Department of Chemistry, Université de Montréal, Montréal, Quebec H3C 3J7, Canada., Bonneil E; Institute for Research in Immunology and Cancer (IRIC)Université de Montréal, Montréal, Quebec H3C 3J7, Canada., Bailey DJ; ThermoFisher Scientific, San Jose, California 95134, United States., Prasad S; ThermoFisher Scientific, San Jose, California 95134, United States., Belford M; ThermoFisher Scientific, San Jose, California 95134, United States., Rochon J; Phytronix Technologies, Québec, Quebec G1P 2J7, Canada., Picard P; Phytronix Technologies, Québec, Quebec G1P 2J7, Canada., Lacoursière J; Phytronix Technologies, Québec, Quebec G1P 2J7, Canada., Dunyach JJ; ThermoFisher Scientific, San Jose, California 95134, United States., Thibault P; Institute for Research in Immunology and Cancer (IRIC)Université de Montréal, Montréal, Quebec H3C 3J7, Canada.; Department of Chemistry, Université de Montréal, Montréal, Quebec H3C 3J7, Canada.
Jazyk:	angličtina
Zdroj:	Analytical chemistry [Anal Chem] 2021 Jul 20; Vol. 93 (28), pp. 9817-9825. Date of Electronic Publication: 2021 Jul 02.
DOI:	10.1021/acs.analchem.1c01376
Abstrakt:	High-field asymmetric waveform ion mobility spectrometry (FAIMS) has gained popularity in the proteomics field for its capability to improve mass spectrometry sensitivity and to decrease peptide co-fragmentation. The recent implementation of FAIMS on Tribrid Orbitrap instruments enhanced proteome coverage and increased the precision of quantitative measurements. However, the FAIMS interface has not been available on older generation Orbitrap mass spectrometers such as the Q-Exactive. Here, we report the integration of the FAIMS Pro device with embedded electrical and gas connections to a Q-Exactive HF mass spectrometer. Proteomic experiments performed on HeLa tryptic digests with the modified mass spectrometer improved signal to noise and reduced interfering ions, resulting in an increase of 42% in peptide identification. FAIMS was also combined with segmented ion fractionation where 100 m / z windows were obtained in turn to further increase the depth of proteome analysis by reducing the proportion of chimeric MS/MS spectra from 50 to 27%. We also demonstrate the application of FAIMS to improve quantitative measurements when using isobaric peptide labeling. FAIMS experiments performed on a two-proteome model revealed that FAIMS Pro provided a 65% improvement in quantification accuracy compared to conventional LC-MS/MS experiments.
Databáze:	MEDLINE
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