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Description
Electron-transfer dissociation (ETD) is a well-established technique, where fragmentation is caused in cationic precursors by electron transfer from a donor molecule. While ETD/EThcD is already present in the family of Orbitrap Tribrid mass spectrometers, using Fluoranthene as an electron donor, this functionality is missing on the Orbitrap Hybrid mass spectrometers. With the Orbitrap Excedion Pro mass spectrometers, we introduce a Orbitrap Hybrid mass spectrometer with ETD and EThcD capabilities. Using the same electron donor as the Orbitrap Tribrid systems, ETD /EThcD reactions can be performed in the ion routing multipole (IRM).
Here, we want to present the performance of the ETD / EThcD functionality of the Orbitrap Excedion Pro mass spectrometer on the most common user cases of this fragmentation technique:
1) Detection of post translational modifications (PTM): Higher-energy Collision Dissociation (HCD) fragmentation is usually too harsh and causes the dissociation of the PTM from the connected peptide in the instrument. This can be avoided using ETD/EThcD as fragmentation technique
2) Differentiation between isomeric amino acids: ETD/EThcD can cause alternate fragmentations in isomeric amino acids like Asp/Iso-Asp and Leu/Ile. These alternate fragments (e.g. w-ions) can be used to differentiate between isomeric amino acids.
3) Top/Middle down analysis of proteins: Using ETD and especially EThcD fragmentation, a much higher sequence coverage can be achieved for the Top/Middle down analysis of proteins compared to HCD fragmentation
4) Analysis of disulfide bonds: Disulfide bonds can be cleaved in the mass spectrometer using ETD/EThcD fragmentation. This allows disulfide linked peptides to be analyzed as MS cleavable crosslinks.
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