Online 2D-LC for Complex N-Glycan Analysis from Biopharmaceuticals

EPO has a complex glycosylation pattern with differently branched and charged glycans. A combination of hydrophilic interaction chromatography (HILIC) with weak anion exchange chromatography (WAX) enables highly orthogonal separation. Comprehensive 2D-LC analysis with HILIC in the first and WAX in the second dimension provides high resolution 2D chromatography together with simultaneous charge profiling. Meanwhile, multiple heart-cutting 2D-LC analysis combining WAX and HILIC separation provides a flexible alternative whereby the user can select multiple peaks to be analyzed in the second dimension and, moreover, run longer gradients in the second dimension.