Projects per year
Abstract
High-field asymmetric waveform ion mobility spectrometry (FAIMS) enables the separation of ions on the basis of their differential mobility in an asymmetric oscillating electric field. We, and others, have previously demonstrated the benefits of FAIMS for the analysis of peptides and denatured proteins. To date, FAIMS has not been integrated with native mass spectrometry of folded proteins and protein complexes, largely due to concerns over the heating effects associated with the high electric fields employed. Here, we demonstrate the newly introduced cylindrical FAIMS Pro device coupled with an Orbitrap Eclipse enables analysis of intact protein assemblies up to 147 kDa. No evidence for dissociation was detected suggesting that any field heating is insufficient to disrupt the noncovalent interactions governing these assemblies. Moreover, the FAIMS device was integrated into native liquid extraction surface analysis (LESA) MS of protein assemblies directly from thin tissue sections. Intact tetrameric hemoglobin (64 kDa) and trimeric reactive intermediate deiminase A (RidA, 43 kDa) were detected. Improvements in signal-to-noise of between 1.5× and 12× were observed for these protein assemblies on integration of FAIMS.
Original language | English |
---|---|
Pages (from-to) | 6811-6816 |
Number of pages | 6 |
Journal | Analytical Chemistry |
Volume | 92 |
Issue number | 10 |
Early online date | 28 Apr 2020 |
DOIs | |
Publication status | Published - 19 May 2020 |
ASJC Scopus subject areas
- Analytical Chemistry
Fingerprint
Dive into the research topics of 'High-field asymmetric waveform ion mobility spectrometry and native mass spectrometry: analysis of intact protein assemblies and protein complexes'. Together they form a unique fingerprint.Projects
- 3 Finished
-
A new mass spectrometer for structural proteomics and protein imaging
Thomas, C., Grant, M., Lovering, A., Alderwick, L., Tomlinson, M., Winn, P., Knowles, T., Cooper, H., Styles, I., Gibbs, D., Huber, D., Bhatt, A. & Leney, A.
Biotechnology & Biological Sciences Research Council
1/07/19 → 30/06/20
Project: Research
-
NAMS: Native ambient mass spectrometry
Engineering & Physical Science Research Council
1/06/19 → 30/11/23
Project: Research
-
A new tool to support drug discovery: Native LESA mass spectrometry (NESA)
Engineering & Physical Science Research Council
1/04/18 → 31/03/21
Project: Research Councils