Expert Insight: Webinar Highlights: Online 2D-LC Analysis of Complex N-Glycans in Biopharmaceuticals

Read the highlights from this SelectScience® webinar

13 Jun 2016


Read the highlights from this SelectScience® webinar

 

Complex glycosylation patterns with differently branched and charged glycans can be resolved with a combination of hydrophilic interaction chromatography (HILIC) and weak anion exchange chromatography (WAX), enabling highly orthogonal separation. Comprehensive 2D-LC analysis with HILIC in the first and WAX in the second dimension facilitates high-resolution 2D chromatography, together with simultaneous charge profiling.

In this webinar, Sonja Schneider, Application Scientist at Agilent Technologies, discussed different online 2D-LC concepts for the high-resolution analysis of complex N-linked glycans in biopharmaceuticals using 2D-LC with fluorescence and MS detection. Read on for the highlights of the Q&A session, or watch the webinar here.

 

Q: Was the amount of ammonium formate in the salt gradient in the second dimension problematic for the MS sensitivity?

A: No, because ammonium formate is an MS-tolerable salt. I obtained good MS and MS/MS spectra after the second dimension.

 

Q: What polarity did you use for the MS measurements?

A: For 2-AB labelled samples, I was using negative polarity, but for the new instant PC-labelled samples positive polarity is recommended as the high sensitivity found in this label is mostly found in positive polarity.

 

Q: Have you tried glycan analysis without labelling? Why not?

A: No - I haven’t tried unlabeled glycans, because I was mostly looking for a FLD solution.

 

 

Watch the presentation here or learn more about biopharmaceuticals with our special feature.