Webinar Highlights: Online 2D-LC Analysis of Complex N-Glycans in Biopharmaceuticals
Read the highlights from this SelectScience® webinar
12 Jun 2016Read 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.
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