Enhancing bottom-up and top-down proteomic measurements with ion mobility separations

Area of Science:

• Proteomics and Analytical Chemistry
• Advancements in Mass Spectrometry and Separation Science

Background:

• High-throughput, sensitive, and structurally informative proteomic measurements are crucial for complex mixture characterization and targeted studies.
• Liquid chromatography coupled with mass spectrometry (LC-MS) has enabled the analysis of thousands of proteins but can be enhanced with additional separation.
• Ion mobility separations (IMS) offer rapid structural information and improved component resolution, complementing existing proteomic workflows.

Purpose of the Study:

• To demonstrate the benefits of incorporating ion mobility spectrometry into mass spectrometry (IMS-MS) for proteomic measurements.
• To illustrate how LC-IMS-MS enhances sensitivity, throughput, and structural information in proteomic analyses.
• To showcase applications in both bottom-up and top-down proteomics.

Main Methods:

• Integration of ion mobility spectrometry (IMS) as a separation dimension within LC-MS workflows.
• Utilizing advanced ion transmission and data acquisition techniques for IMS-MS and LC-IMS-MS.
• Application of these methods to both bottom-up and top-down proteomic measurement strategies.

Main Results:

• Demonstrated increased sensitivity in proteomic measurements through the application of IMS-MS and LC-IMS-MS.
• Achieved higher throughput in proteomic analyses by leveraging the speed of ion mobility separations.
• Obtained enhanced structural information of proteins and peptides, improving the depth of proteomic characterization.

Conclusions:

• Ion mobility spectrometry is a valuable technology for enhancing LC-MS-based proteomic measurements.
• The incorporation of IMS significantly improves sensitivity, throughput, and the acquisition of structural data in proteomics.
• LC-IMS-MS represents a powerful approach for comprehensive proteomic studies, applicable to diverse measurement strategies.