The ExD WK-150 Option extends the capability of Waters SELECT SERIES™ Cyclic IMS systems to include electron-based dissociation. Fast electron capture dissociation (ECD) coupled with high-resolution cIM enhances IMSn analytical workflows investigating the structure and heterogeneity of complex biomolecules.
The ExD WK-150 Option makes electron-based fragmentation methods accessible and highly affordable for pharmaceutical, clinical, consumer product, and life science protein research laboratories.
Increase sequence coverage of larger peptides and intact proteins beyond the limits of CID alone.
Since ECD and CID produce complementary sequence information, the combination of both methods increases confidence in results.
ECD fragmentation products unique to isoaspartate clearly differentiate it from the non-isomerized form. Isoaspartate is implicated in age-related protein inactivation and aggregation, and reduced efficacy of protein therapeutics.
Side chain fragments (w-ions) produced by ECD can be used to distinguish isobaric amino acids.
The ExD Cell can operate in either ECD or transmission-only mode.
Our ExD filament design features a plug-in cassette with exchangeable filament insert for quick and easy replacement.
Add ExD to your instrument purchase order to capitalize on the unique IM capability of the SELECT SERIES™ Cyclic IMS platform.
The ExD cell sits either before or after the cyclic ion guide, providing optional ECD fragmentation of either isolated precursor ions or separated conformers respectively.
The ExD Controller is built into the mass spectrometer.
ECD is a “gentle” fragmentation method that preserves labile PTMs – phosphorylation, glycosylation – on fragment ions. With the ExD cell installed, use ECD to map their precise location on peptides and proteins.
Selective dissociation of disulfide bonds by ECD enables their localization without prior sample reduction.
The ExD WK-150 Option includes:
Beckman, J. S.; Voinov, V. G.; Hewitt, D.; Wildgoose, J.; Williams, J. P.; Brown, J. M.; Langridge, J. I.; Cooper-Shepherd, D. A. Implementation and evaluation of electron capture dissociation (ECD) on a cyclic IMS enabled mass spectrometer. Poster presented at: 68th Annual ASMS Conference on Mass Spectrometry and Allied Topics 2020 Reboot. 2020. Online.
e-MSion uses “ExD” to refer to the broad range of electron energies the ExD cell can produce. The ExD cell is capable of both low-energy (below 3 eV) electron capture dissociation (ECD) and high-energy (3-20 eV) electron-induced dissociation (EID).
In the literature, “ExD” refers to a family of electron-based fragmentation technologies including ECD, EID, electron transfer dissociation (ETD), electron detachment dissociation (EDD), and negative ion ECD (niECD). EID is also sometimes split into hot ECD (HECD) and electron-impact excitation of ions from organics (EIEIO), depending on the electron energy used.
See Qi and Volmer. 2017. Electron-based Fragmentation Methods in Mass Spectrometry: An Overview. Mass Spectrometry Reviews 36, 4-15.
e-MSion currently offers a commercial ExD solution for Agilent LC/Q-TOF and Waters SELECT SERIES™ Cyclic IMS mass spectrometers.
Contact us if you are interested in becoming an early adopter of ExD for a mass spectrometer belonging to the Thermo Fisher Q Exactive Orbitrap™ series or the Waters SYNAPT™ G2 series.
Please contact e-MSion directly for service and support inquiries.
Including time for unpacking, venting, pump down, and performance verification, a field service engineer will typically require 2 days to complete the installation process. Hardware modification takes about 30 minutes.
Please contact us to learn more about installation requirements.
The ExD cell operates on a microsecond timescale. Its high speed and flow-through design means that the instrument duty cycle is unaffected by installation of the cell.
The ExD cell electron source is a consumable filament, which requires replacement after burning out. Several features in the ExD Software are designed to extend the filament lifespan by protecting it from rapid heat changes.
Filament replacement involves swapping out “plug-in” filament cassettes inside the cell, which minimizes the time spent with the instrument vented and the subsequent pump down time needed before the instrument is operational again. We provide users with instructions for replacing the filament in our product documentation.
Unlike ETD, ECD does not use any reagent. This combined with its distance from the source means the ExD cell does not require regular cleaning.
Use the ExD Software to change ExD cell settings. The same settings for ECD will work on most analytes with minimal adjustment. See Technology for a description of ExD cell operation.
Currently, the ExD AQ-250 Option for Agilent LC/Q-TOF offers an autotune feature for simplified use. Autotuning for our other products is under development.
Due to its flow-through design, the ExD cell operates at a speed compatible with HPLC, UPLC, CE, and IM separation methods.
The ExD cell is designed to fragment polypeptides, but can also be tuned to fragment glycans and lipids.
ECD efficiency theoretically increases with the square of ion charge, making the ExD cell more efficient at fragmenting large, multi-charged proteins. In practice, noncovalent interactions limit the ability of ECD alone to dissociate folded proteins larger than ~30kDa. Denaturing conditions and/or supplemental vibrational excitation can be used to increase ECD efficiency of larger proteins.
On the other end of the spectrum, the ExD cell is capable of fragmenting short peptides, albeit with lower efficiency. For ECD, a minimum precursor charge of 2+ is required to detect product ions because of the charge neutralization that occurs with electron capture.
The ExD cell can also be tuned for EID to target singly charged precursors and non-peptidic samples. This technique is currently only useful for applications where sample quantity is not limited. Development to increase EID efficiency is ongoing.
At some point, you or your service engineer may want to temporarily revert the instrument to its default hardware configuration. The “swap-out” design of the ExD cell makes this possible, since all original parts are preserved during installation.