pr3c00027_si_004.xlsx (26.83 kB)
High-Field Asymmetric Waveform Ion Mobility Spectrometry: Practical Alternative for Cardiac Proteome Sample Processing
datasetposted on 2023-04-11, 23:33 authored by Lizhuo Ai, Aleksandra Binek, Simion Kreimer, Matthew Ayres, Aleksandr Stotland, Jennifer E. Van Eyk
Heart tissue sample preparation for mass spectrometry (MS) analysis that includes prefractionation reduces the cellular protein dynamic range and increases the relative abundance of nonsarcomeric proteins. We previously described “IN-Sequence” (IN-Seq) where heart tissue lysate is sequentially partitioned into three subcellular fractions to increase the proteome coverage more 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. The FAIMS approach substantially reduces manual sample handling, significantly shortens the MS instrument processing time, and produces unique protein identification and quantification approximating the commonly used IN-Seq method in less time.
three subcellular fractionspreviously described “includes prefractionation reducesheart tissue lysatesignificantly shortenssequentially partitionedsequence ”relative abundancequantification approximatingproteome coveragepractical alternativephase fractionationnonsarcomeric proteinsmass spectrometryless timecommonly used