biostudies-literatureJager SDutch Research Council (NWO)2497https://www.ebi.ac.uk/biostudies/studies/S-EPMC11906852biostudies-literatureUnknown16(1)Recently, a conceptually new mass analyzer was introduced by pairing a quadrupole Orbitrap mass spectrometer with an asymmetric track lossless (Astral™) analyzer. This system provides >200 Hz MS/MS scanning speed, high resolving power, sensitivity, and mass accuracy. Due to its speed, the instrument allows for a narrow-window data-independent acquisition (nDIA) strategy, representing a new technical milestone in peptide-centric proteomics. However, this new system may also be applied to other complex and clinically important proteomes, such as the human plasma N-glycoproteome. Here, we evaluate the Orbitrap Astral mass spectrometer for the in-depth analysis of the plasma N-glycoproteome and pioneer a dedicated nDIA workflow, termed "nGlycoDIA", on glycopeptide enriched and crude plasma. ThNature communicationsIn-depth plasma N-glycoproteome profiling using narrow-window data-independent acquisition on the Orbitrap Astral mass spectrometer.PMC11906852184.034.019SPI.2017.028Heck AJRDamoc EZeller MReiding KRMakarov AAPashkova AJager SSchulte DfalseIn-depth plasma N-glycoproteome profiling using narrow-window data-independent acquisition on the Orbitrap Astral mass spectrometer.Recently, a conceptually new mass analyzer was introduced by pairing a quadrupole Orbitrap mass spectrometer with an asymmetric track lossless (Astral™) analyzer. This system provides >200 Hz MS/MS scanning speed, high resolving power, sensitivity, and mass accuracy. Due to its speed, the instrument allows for a narrow-window data-independent acquisition (nDIA) strategy, representing a new technical milestone in peptide-centric proteomics. However, this new system may also be applied to other complex and clinically important proteomes, such as the human plasma N-glycoproteome. Here, we evaluate the Orbitrap Astral mass spectrometer for the in-depth analysis of the plasma N-glycoproteome and pioneer a dedicated nDIA workflow, termed "nGlycoDIA", on glycopeptide enriched and crude plasma. Th2025-01-01T00:00:00Z2025 Mar2026-06-01T14:37:19.469Z2025-04-04T00:06:34.004ZS-EPMC119068524008247410.1038/s41467-025-57916-1