Improvements in glycoproteomics through architecture changes to the Orbitrap Tribrid MS platform DOI Creative Commons
Tim S. Veth, Emmajay Sutherland, Ruby Zhang

et al.

Published: Aug. 29, 2024

Hardware changes introduced on the Orbitrap Ascend MultiOmics Tribrid MS include dual ion routing multipoles (IRMs) that can enable parallelized accumulation, dissociation, and mass analysis of three separate populations. The balance between these instrument functions is especially important in glycoproteomics, where complexities glycopeptide fragmentation necessitate large precursor populations and, consequently, long accumulation times for quality MS/MS spectra. To compound matters further, dissociation methods like electron transfer (ETD) benefit characterization come with overhead also slow down scan acquisition. Here we explored how IRM architecture improve analysis, a focus O-glycopeptide using ETD supplemental collisional activation (EThcD). We found parallelization EThcD – uniquely enabled by increased acquisition speed without sacrificing spectral quality, subsequently increasing number O-glycopeptides identified relative to analyses Eclipse (i.e., previous generation MS). Additionally, systematically evaluated ion-ion reaction energies used understand best utilize time architecture. observed shorter-than-expected minimized c/z•-fragment generation, higher collision generate combinations glycan-retaining glycan-neutral-loss peptide backbone fragments identification. saw improvements N-glycopeptide collision-based faster speeds. Overall, data show architectural platform glycoproteomic experiments parallelizing minimize sensitivity.

Language: Английский

Improvements in glycoproteomics through architecture changes to the Orbitrap Tribrid MS platform DOI Creative Commons
Tim S. Veth, Emmajay Sutherland, Ruby Zhang

et al.

Published: Aug. 29, 2024

Hardware changes introduced on the Orbitrap Ascend MultiOmics Tribrid MS include dual ion routing multipoles (IRMs) that can enable parallelized accumulation, dissociation, and mass analysis of three separate populations. The balance between these instrument functions is especially important in glycoproteomics, where complexities glycopeptide fragmentation necessitate large precursor populations and, consequently, long accumulation times for quality MS/MS spectra. To compound matters further, dissociation methods like electron transfer (ETD) benefit characterization come with overhead also slow down scan acquisition. Here we explored how IRM architecture improve analysis, a focus O-glycopeptide using ETD supplemental collisional activation (EThcD). We found parallelization EThcD – uniquely enabled by increased acquisition speed without sacrificing spectral quality, subsequently increasing number O-glycopeptides identified relative to analyses Eclipse (i.e., previous generation MS). Additionally, systematically evaluated ion-ion reaction energies used understand best utilize time architecture. observed shorter-than-expected minimized c/z•-fragment generation, higher collision generate combinations glycan-retaining glycan-neutral-loss peptide backbone fragments identification. saw improvements N-glycopeptide collision-based faster speeds. Overall, data show architectural platform glycoproteomic experiments parallelizing minimize sensitivity.

Language: Английский

Citations

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