Ion mobility-mass spectrometry is an emerging gas-phase technology that separates ions, based on their collision cross-section and mass. The method provides three-dimensional information on the overall topology and shape of protein complexes. Here, we outline a basic procedure for instrument setting and optimization, calibration of drift times, and data interpretation.
Total time
~4–6 hours per sample (instrument setup, acquisition, and data processing)
Steps
1
Understand ion mobility-mass spectrometry principles and applications
Learn the fundamental concepts of gas-phase ion separation by collision cross-section and mass, and how this technique provides structural topology information for protein complexes.
▶ 00:46
2
Acquire ion mobility-mass spectrometry spectrum
Configure instrument parameters and perform the complete data acquisition workflow to generate ion mobility and mass spectrometry data from protein complex samples.
▶ 01:15
3
Screen and optimize experimental conditions
Systematically test and refine instrument settings and sample preparation to maximize data quality and signal intensity.
▶ 07:49
4
Correlate drift time values with cross-sectional areas
Calibrate the instrument using reference standards and convert experimental drift time measurements into collision cross-section values for structural interpretation.
▶ 09:45
5
Interpret and analyze representative results
Review exemplar datasets to understand output formats, validate structural assignments, and extract biologically meaningful topological information from protein complexes.
▶ 13:26