Detailed energy dependence investigations were carried out on AGP digest using 2.2 pmol glycoprotein per run and on the mixture of AGP, fetuin and transferrin digests injecting 2.2 pmol from all glycoproteins in each run. Stepped CE settings were applied with 80% of the time allocated to the higher energy component and the low CE/high CE ratio was set to 0.5. The CE was systematically varied from 6.25% to 175% of the Hinneburg et al.'s setting in steps of 6.25% resulting in 27 different nano-LC-MS/MS runs. Experiments were performed with the use of three inclusion lists, based on DDA measurements taken with Hinneburg et al.'s CE method. Two lists for the mixture (3GP - SPLv5b, SPLv6) and one list for AGP (AGP - SPLv7) were created. The higher component of the collision energy is marked by a 3,4,5 digit number after the SPL designation, containing 0,1,2 decimal digits, respectively (eg "100"->100%, "08125"-->81.25%).
[doi:10.25345/C51834621]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: N-glycosylation ; collision energy ; optimization ; transfer ; monoclonal antibody
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Principal Investigators: (in alphabetical order) |
Agnes Revesz, Research Centre for Natural Sciences, Budapest, Hungary |
| Submitting User: | reveszagnes |
Hevér H, Nagy K, Xue A, Sugár S, Komka K, Vékey K, Drahos L, Révész Á.
Diversity Matters: Optimal Collision Energies for Tandem Mass Spectrometric Analysis of a Large Set of N-Glycopeptides.
J Proteome Res. 2022 Nov 4;21(11):2743-2753. Epub 2022 Oct 6.
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