Process Step Action
Affinity Capture & Cleanup Protein sample is captured, and its matrix components are removed.
Release of Glycan Enzyme PNGase F is pushed into the affinity column where purified protein resides and glycans are released into interstitial space of the column. Released free glycans are removed from the affinity column.
Glycan Labeling The release free glycans and glycan labeling reagent are mixed, degassed, and heated inside the reaction coil where the labeling reaction occurs.
Cooling and Enrichment of the labeled glycans The reaction mixture is delivered to the cooling coil and labeled glycans are captured and enriched in the PGC column. Excess reagent is removed as the flow-through.
Prepare for LC Injection Labeled glycans are eluted from the PGC column. The organic content of the PGC eluted sample is adjusted in the mixing chamber before loading to the LC sample loop and initiate HILIC-LC method for glycan quantitation.