High-Field Asymmetric Waveform Ion Mobility Spectrometry: Practical Alternative for Cardiac Proteome Sample...

Area of Science:

• Proteomics
• Biochemistry
• Analytical Chemistry

Background:

• Mass spectrometry (MS) analysis of heart tissue benefits from prefractionation to reduce protein dynamic range and increase nonsarcomeric protein abundance.
• The previously developed IN-Sequence (IN-Seq) method sequentially partitions heart tissue lysate into subcellular fractions for enhanced proteome coverage.

Purpose of the Study:

• To adapt high-field asymmetric ion mobility spectrometry (FAIMS) coupled with MS for heart tissue sample preparation.
• To establish a simplified, one-step sample preparation method utilizing gas-phase fractionation.
• To compare the efficiency and proteome coverage of the FAIMS approach with the established IN-Seq method.

Main Methods:

• Adaptation of high-field asymmetric ion mobility spectrometry (FAIMS) for coupling with mass spectrometry.
• Development of a one-step sample preparation protocol involving gas-phase fractionation.
• Sequential partitioning of heart tissue lysate into subcellular fractions (IN-Seq) as a comparative method.

Main Results:

• The FAIMS approach significantly reduces manual sample handling and shortens MS instrument processing time.
• FAIMS-based sample preparation yields unique protein identification and quantification comparable to the IN-Seq method.
• This novel method achieves similar proteome coverage to IN-Seq in a substantially shorter timeframe.

Conclusions:

• High-field asymmetric ion mobility spectrometry (FAIMS) coupled with MS offers an efficient alternative for heart tissue proteomic analysis.
• The one-step FAIMS sample preparation simplifies workflows, reduces hands-on time, and accelerates MS data acquisition.
• This method provides a valuable tool for comprehensive heart tissue proteome analysis, approximating the depth of IN-Seq with improved efficiency.