Poster Presentation AUS-oMicS 2025

Enhancing glycopeptide characterization using nanoflow LC separation and electron activated dissociation fragmentation (121173)

Jeremy JP Potriquet 1 , Edward EK Kerr 2 , Amanda AN Nouwens 3 , Susan SB Briscoe 2 , Tony TV Vuocolo 2
  1. SCIEX, Mt Waverley, VIC, Australia
  2. CSIRO, Brisbane
  3. UQ, Brisbane

This technical note describes a workflow for a more comprehensive characterization of glycoproteins using nanoflow liquid chromatography (LC) separation along with collision-induced dissociation (CID) and electron activated dissociation (EAD) fragmentation on the ZenoTOF 7600 system. LC separation using IonOpticks Aurora Elite SX nanoflow columns resulted in 3-5x gains in glycopeptide identifications compared to microflow LC separation. The high degree of MS/MS sensitivity with Zeno trap pulsing on the ZenoTOF 7600 system, in combination with nanoflow LC separation, enables the identification of glycosylated tryptic peptides with a data-dependent acquisition (DDA) approach using CID fragmentation. EAD fragmentation produces complementary peptide fragment ions, allowing for a greater depth of characterization through unambiguous assignment of the types and sites of glycosylation post-translational modifications