Site-specific glycoprofiling of pharmaceutical compounds employing quantitative glycopeptide enrichment followed by complementary HILIC-and RP-LC-MS analysis

摘要

The ZIC~(~R) Glycocapture Resin allows for specific enrichment of N- and O-linked glycopeptides without preference for particular glycan structures or loss of information. The use of proteinase K or a combination of two specific proteases (e.g. trypsin +Glu-C) facilitates efficient O-glycopeptide enrichment using the same procedure. RP-18 and ZIC~(~R)-HILIC chromatography are orthogonal and complementary for separation of glycopeptides. The use of both stationary phases in 1- and 2-dimensional LC-MS setups results in the detection of low abundant glycoforms, which, can not be identified if only one stationary phase is employed.