Poster Presentation 28th Annual Lorne Proteomics Symposium 2023

A novel fragmentation technology allows for in-depth glycopeptide characterization in glycoproteins (#162)

Jenny Albanese 1 , Zoe Zhang 1 , Jeremy Potriquet 2
  1. SCIEX, South San Francisco, California, USA
  2. SCIEX, Mt Waverley, VIC, Australia

Improving the depth of characterization of glycoprotein is essential in the field of biotherapeutics and mAbs analysis.
The newly developed electron activated dissociation (EAD) approach allows to tune electron energy to produce different fragmentation patterns of the electron-based dissociation (ExD) family.

In this study monoclonal antibodies adalimumab, rituximab as well as an antibody-drug-conjugated sample (ado- trastuzumab emtansine) were digested into peptides and acquired on the ZenoTOF 7600 using fast data-dependent acquisition (DDA) combined with EAD alternative fragmentation.

This novel fragmentation technology resulted in rich fragmentation with predominantly peptide backbone fragments leaving the glycan intact on the glycosylation site, enabling localization and providing a higher level of structural information for glycopeptide characterization.
Thanks to the Zeno trap the detection level of the diagnostic fragments was increased by 5 to 10-fold

The data presented here demonstrate the advantage for glycoproteins using this novel fragmentation cell over traditional collision-induced dissociation (CID) used for peptide mapping, which does not allow for consistent identification and localization of glycans on peptides. Combining increased detection of fragments using a mechanism for increased duty cycle on a Q-TOF instrument (the Zeno trap) enables higher confidence in data assignment, making the Zeno EAD combination ideal for in-depth analysis of glycopeptides.
We are also demonstrating the power of the SCIEX OS software and automated data interpretation with the new Biologics Explorer software offering a streamlined characterization workflow.