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Integration of FAIMS on a Q-Exactive HF mass spectrometer

ABSTRACT: 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 150 m/z windows were analyzed 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.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Eric Bonneil

LAB HEAD: Pierre Thibault

PROVIDER: PXD019848 | Pride | 2021-02-16

REPOSITORIES: Pride

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