High-Field Asymmetric Waveform Ion Mobility Spectrometry: Practical Alternative for Cardiac Proteome Sample Processing.
Publication Year:
2023
PubMed ID:
37040897
Funding Grants:
Public Summary:
Preparing heart tissue for protein analysis by mass spectrometry usually involves breaking it down into parts to better detect less common proteins. A method called IN-Seq does this by separating the tissue into three parts to get more detailed results. This study introduces a new approach using a technology called FAIMS that simplifies the process into one step, reduces the time spent preparing samples and running the analysis, and still finds almost as many proteins as the older method. This makes protein analysis faster and easier without losing much detail.
Scientific Abstract:
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.