Enhancing the Peak Capacity of High Molecular Weight N-Glycan HILIC Separations with a Wide-Pore Amide Bonded Stationary Phase

Library Number:
APNT134848086
Part Number:
720005381EN
Author(s):
Matthew A. Lauber and Stephan M. Koza
Source:
Waters
Content Type:
Application Notes
Content Subtype:
Technical Notes
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High molecular weight, tri- and tetra-antennary N-glycans are highly branched structures that adopt relatively large radii of hydration in solution. To achieve optimal HILIC separations of these large structures, we propose a column with a wide-pore amide bonded stationary phase, a glycoprotein BEH amide, 300Å, 1.7 μm column. For large glycan species, this column provides increases in peak capacity over a conventional pore diameter column of approximately 17%. Improved resolving power is particularly useful in this separation space as it is typified by highly complex glycan profiles. Most notably, these improvements in resolution should be of significant utility in the characterization and routine monitoring of biopharmaceuticals that are expressed with large, highly complicated N-glycan structures, such as coagulation Factor IX, erythropoietin, and darbepoetin.


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