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bioRxiv · 10.1101/2022.12.28.522124

High Field Asymmetric Waveform Ion Mobility Spectrometry- Mass Spectrometry to Enhance Cardiac Muscle Proteome Coverage

Ai, L.·Binek, A.·Kreimer, S.·Ayres, M.·Stotland, A.·Van Eyk, J. E.
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Abstract

Heart tissue sample preparation for mass spectrometry (MS) analysis that includes pre-fractionation reduces the cellular protein dynamic range and increases the relative abundance of non-sarcomeric proteins. We previously described "IN-Sequence" (IN-Seq) where heart tissue lysate is sequentially partitioned into three subcellular fractions to increase the proteome coverage than a single direct tissue analysis by mass spectrometry. Here, we report an adaptation of the high-field asymmetric ion mobility spectrometry (FAIMS) coupled to mass spectrometry, and the establishment of a simple one step sample preparation coupled with gas-phase fractionation. FAIMS approach substantially reduces manual sample handling, significantly shortens MS instrument processing time, and produces unique protein identification and quantification approximating the commonly used IN-Seq method in for less time requirement.

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Ai, L., Binek, A., Kreimer, S., Ayres, M., Stotland, A., Van Eyk, J. E.. 2022-12-29. High Field Asymmetric Waveform Ion Mobility Spectrometry- Mass Spectrometry to Enhance Cardiac Muscle Proteome Coverage. https://doi.org/10.1101/2022.12.28.522124

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