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. |