Broadband ion mobility deconvolution for rapid analysis of complex mixtures

Area of Science:

• Analytical Chemistry
• Biochemistry
• Proteomics

Background:

• High-resolution ion mobility (IM) is crucial for analyzing complex mixtures in high-throughput IM mass spectrometry (IM-MS).
• Previous methods used automated ion mobility deconvolution (AIMD) software with m/z-isolation for pure component IM-MS data extraction.
• Ion isolation steps limited experimental throughput and applicability.

Purpose of the Study:

• To develop a broadband IM-MS deconvolution strategy eliminating the need for m/z-isolation.
• To enhance the applicability of AIMD for high-throughput bottom-up proteomics by reducing experimental run times.
• To demonstrate the efficacy of broadband deconvolution for resolving isomeric species and complex biological samples.

Main Methods:

• Implementation of a broadband IM-MS deconvolution strategy.
• Removal of the m/z-isolation requirement for deconvolution of IM unresolved peaks.
• Application of the method to simulated complex mixtures of isomeric hexapeptides and trisaccharides.
• Analysis of a rat brain tissue proteolytic digest using IM-assisted data-independent analysis (DIA/HDMSE).

Main Results:

• Successful deconvolution of IM unresolved isomeric hexapeptides and trisaccharides using broadband IM-MS.
• Demonstrated improvement in high-throughput bottom-up characterization of a complex biological sample (rat brain tissue digest).
• Achieved deconvolution of pure component IM and MS data from an IM-assisted DIA/HDMSE dataset, a novel accomplishment.

Conclusions:

• Broadband IM-MS deconvolution effectively resolves IM unresolved peaks without m/z-isolation.
• Eliminating ion isolation significantly reduces experimental time, expanding AIMD's utility in high-throughput proteomics.
• This study presents the first successful deconvolution of IM and MS data from IM-assisted DIA/HDMSE, advancing proteomic analysis capabilities.