11
Native glycan profiles are significantly different from the
profile of fluorescently labeled glycans, especially for
glycans containing multiple sialic acids (Figure 3).
However, labeled glycans require smaller amounts
(10 times) of samples for MS analysis as compared to
native glycans. Thus, the GlycanPac AXH-1 column is
useful for the analysis of biologically relevant glycans
including glycans from antibodies, either labeled or
native, by LC-fluorescence or LC-MS methods. If the
amount of the sample is not extremely limited, analysis
of unlabeled glycans using the GlycanPac AXH-1 is
highly feasible.
Conclusion
• A fully integrated workflow for structural
characterization of native and fluorescently labeled
N
-glycans released from proteins was demonstrated
successfully.
• Novel GlycanPac AXH-1 column demonstrated
excellent separation of released N-glycans especially
forsilalylated species. It allowed for their sensitive
detection by the Q Exactive mass spectrometer and
identification by SimGlycan software.
• This LC-MS integrated technology is also useful for the
separation and structural characterization of reduced
O
-linked glycans from proteins, mucins, and the
analysis of charged and neutral glycosylaminoglycans
and glycolipids.
References
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3.Guidance for Industry, Scientific Considerations in
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