Open AccessAdvanced Strategies for Proton-Transfer Reactions Coupled with Parallel Ion Parking on a 21 T FT-ICR MS for Intact Protein Analysis
Author(s) -
Chad R. Weisbrod,
Lissa C. Anderson,
Christopher L. Hendrickson,
Leah V. Schaffer,
Michael R. Shortreed,
Lloyd M. Smith,
Jeffrey Shabanowitz,
Donald F. Hunt
Publication year - 2021
Publication title -
analytical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.117
H-Index - 332
eISSN - 1520-6882
pISSN - 0003-2700
DOI - 10.1021/acs.analchem.1c00847
Subject(s) - chemistry , ion , mass spectrometry , analyte , analytical chemistry (journal) , tandem mass spectrometry , reagent , proton , tandem , chromatography , physics , materials science , organic chemistry , quantum mechanics , composite material
Proton-transfer reactions (PTRs) have emerged as a powerful tool for the study of intact proteins. When coupled with m / z -selective kinetic excitation, such as parallel ion parking (PIP), one can exert exquisite control over rates of reaction with a high degree of specificity. This allows one to "concentrate", in the gas phase, nearly all the signals from an intact protein charge state envelope into a single charge state, improving the signal-to-noise ratio (S/N) by 10× or more. While this approach has been previously reported, here we show that implementing these technologies on a 21 T FT-ICR MS provides a tremendous advantage for intact protein analysis. Advanced strategies for performing PTR with PIP were developed to complement this unique instrument, including subjecting all analyte ions entering the mass spectrometer to PTR and PIP. This experiment, which we call "PTR-MS 1 -PIP", generates a pseudo-MS 1 spectrum derived from ions that are exposed to the PTR reagent and PIP waveforms but have not undergone any prior true mass filtering or ion isolation. The result is an extremely rapid and significant improvement in the spectral S/N of intact proteins. This permits the observation of many more proteoforms and reduces ion injection periods for subsequent tandem mass spectrometry characterization. Additionally, the product ion parking waveform has been optimized to enhance the PTR rate without compromise to the parking efficiency. We demonstrate that this process, called "rapid park", can improve reaction rates by 5-10× and explore critical factors discovered to influence this process. Finally, we demonstrate how coupling PTR-MS 1 and rapid park provides a 10-fold reduction in ion injection time, improving the rate of tandem MS sequencing.