N-Glycosylation Pattern of E2 Glycoprotein from Classical Swine Fever Virus

The extracellular domain of E2 glycoprotein outer surface of the classical swine fever virus was expressed in epithelial kidney pig cells. The N-glycosylation determined by combination of Normal Phase-HPLC, Weak Anion Exchange-HPLC, exoglycosidase digestions and Mass Spectrometry revealed a complex mixture of neutral and monosialylated multiantennary N-glycans with variable number of alpha 1-3-Gal-Gal antennae terminals. The most abundant neutral N-glycan has a composition of HeX(7)HexNAC4dHexl, Negative ion ESI-MS/MS confirmed the presence of the 0-3-Gal-Gal motif on each arm of the fucosylated biantennary N-glycan. The most abundant monosialylated glycan was HeX6HexNAC4d Hex, Neu5Acl, with the sialic acid linked to the terminal fl1-4-Gal-G1cNAc. Sialic acid on the antenna capping position was predominantly of the N-acetyl form.