Work

Deeper Protein Identification by Using FAIMS in Top-down Proteomics

Year: 2021

Type: preprint

Abstract: Field Asymmetric Ion Mobility Spectrometry (FAIMS), when used in proteomics studies, provides superior selectivity, and enables more proteins to be identified by providing additional gas phase separation. Here, we tested the performance of cylindrical FAIMS for the identification and characterization of proteoforms by top-down mass spectrometry of heterogeneous protein mixtures. Combining FAIMS with chromatographic separation resulted in a 62% increase in protein identifications and an 8% increase in proteoform identifications as compared to samples analyzed without FAIMS. This increase was attributable, in part, to improved signal-to-noise for proteoforms with similar retention times. Additionally, our results show that the optimal compensation voltage of any given proteoform was correlated with the molecular weight of the analyte. Collectively these results suggest that the addition of FAIMS can enhance top-down proteomics in both discovery and targeted applications.

Source: ChemRxiv

DOI: 10.26434/chemrxiv.13653578

Authors: Robert Gerbasi, Rafael D. Melani, Susan E. Abbatiello, Michael W. Belford, Romain Huguet, John P. McGee, Dawson Dayhoff, Paul M. Thomas, Neil. L Kelleher

Institutions: Pacific Northwest National Laboratory, Northwestern University, Northeastern University, Thermo Fisher Scientific (United States)

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Awards: National Resource for Translational and Developmental Proteomics, G5091946335

OpenAlex ID: W4242177709

Cites: 29

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Related to: 10