Abstract
Bovine Lf (bLf) possesses five N-glycosylation sites having diverse glycans that alter over lactation and are extensively characterised in milk. However, detailed site-specific characterisation in colostrum is limited. In the present study, the glycan diversity in colostrum bLf from native and commercial preparations was determined by generating N-glycopeptides using trypsin and a combination of trypsin/chymotrypsin followed by RP-UHPLC-Q-TOF MS/MS. The glycan diversity at N300KS site is poorly understood and the analysis revealed the presence of mostly biantennary complex glycans with core fucosylation and differential sialylation. N252NS and N564DT sites were found to possess predominantly high mannose glycans. A range of complex, hybrid and high mannose glycans were determined at N387VT and N495QT sites. Glycan heterogeneity with relative abundance was determined on all glycosylation sites in bLf. In addition, the glycan diversity of lipocalin-2 (N85VT) that might have co-purified in the commercial colostrum bLf sample was also elucidated.
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