4.8 Article

Two-Dimensional FAIMS-IMS Characterization of Peptide Conformers with Resolution Exceeding 1000

Journal

ANALYTICAL CHEMISTRY
Volume 94, Issue 16, Pages 6363-6370

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c00805

Keywords

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Funding

  1. National Natural Science Foundation of China [U20A20121, 61971248]
  2. Key Research and Development Program of Zhejiang [2020C03064]
  3. Natural Science Foundation of Zhejiang Province [LY22F010004]
  4. Ningbo Science and Technology Project [2021S130]
  5. K.C. Wong Magna Fund in Ningbo University [xkzwl1507]

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A high-performance FAIMS-IMS-MS platform was developed for exploring the conformational diversity of bradykinin. The FAIMS-IMS technique can resolve multiple conformers of (BK + H+) ions, and the separations in FAIMS and IMS dimensions are largely orthogonal, resulting in a higher overall resolving power.
A high-performance field asymmetric waveform ion mobility spectrometry (FAIMS)-IMS-MS platform was developed and applied to explore the conformational diversity of the singly and doubly charged bradykinin (BK + H+)(+) and (BK + 2H(+))(2+) ions. With pure N-2 as the FAIMS carrier gas, more than ten conformers of (BK + H+)(+) can be resolved using FAIMS-IMS, as compared to only four conformers resolved using either FAIMS or IMS alone. Interestingly, multiple conformers of (BK + H+)(+) were found to have completely different values of FAIMS compensation voltage (CV), while their IMS drift times were essentially the same, which were also proven experimentally to not result from the structural annealing by the collisional heating in the ion funnel. The separations in the FAIMS and IMS dimensions are substantially orthogonal, and the overall resolving power of two-dimensional FAIMS-IMS separation is largely proportional to the product of the separation resolving powers of FAIMS and IMS. Using a gas mixture of N-2/He to further improve the resolving power of the FAIMS separation, the total resolving powers of the combined FAIMS and IMS separation were estimated to be about 1020 and 1400 for (BK + H+)(+) and (BK + 2H(+))(2+) ions, respectively, which are significantly higher than the resolving power of any ion mobility-based separation techniques demonstrated so far. The combined FAIMS-IMS can thus be a much more powerful technique to explore the structural diversity of biomolecules.

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