Advancing Glycopeptide and Phosphopeptide Analysis with HALO® Penta HILIC

Two recently published studies highlight how researchers are pushing the boundaries of quantitative glycoproteomics and PTM analysis – and both feature the use of HALO® Penta HILIC columns to support reliable, high-resolution separations.

1. Standards-Free Quantitation of Glycopeptides Using Coulometric Mass Spectrometry

This study demonstrates a new approach for absolute quantitation of oxidizable glycopeptides without the need for isotope-labeled standards. Using coulometric mass spectrometry (CMS), the team successfully quantified tyrosine-containing glycopeptides and applied the method to NIST mAb digest samples. HALO® Penta HILIC played a key role in separating N297 glycopeptides prior to CMS analysis, enabling clear resolution of multiple glycoforms and supporting accurate quantitation.

Read more here: https://pubs.acs.org/doi/10.1021/jasms.4c00052 

2. Absolute Quantitation of Phosphopeptides and Glycopeptides Using Coulometric Mass Spectrometry

Building on the CMS framework, this upcoming work extends the technique to phosphopeptides and additional glycopeptide classes. The researchers again leveraged HALO® Penta HILIC to efficiently resolve complex PTM-containing peptides, underscoring the column’s ability to deliver robust retention and sharp peak shapes for highly hydrophilic analytes.

Read more here: https://pubs.acs.org/doi/10.1021/acsmeasuresciau.5c00047 

Why These Papers Matter

Glycopeptide and phosphopeptide quantitation remains a major challenge, especially when standards are unavailable or cost-prohibitive. Both studies show the potential of CMS combined with high-efficiency HILIC separation to simplify workflows and improve quantitative confidence.

HALO® Penta HILIC columns are designed for this type of work, providing:

• Strong retention of hydrophilic and modified peptides
• High efficiency for complex glycoforms
• Reliable performance for PTM-rich samples such as mAbs

To learn more about HALO® Penta HILIC columns, visit https://halocolumns.com/product/halo-90-a-penta-hilic/?gad_source=1&gad_campaignid=13358078299&gbraid=0AAAAADh-UCCjb2uMlTF1qqsWyOmuCa4LI&gclid=CjwKCAiAl-_JBhBjEiwAn3rN7f-JW0h-qHZmJvq5Z-8d-IsBqWuO3wg8yQGCn3e9rMi2ouke9gA4RhoCxKsQAvD_BwE