An innovative method used for the identification of N-glycans on soybean allergen β-conglycinin

β -Conglycinin is one of the major allergens existed in soybean. N -Glycans attached to the β -conglycinin influenced the immunoreactivity and antigen presenting efficiency of β -conglycinin. In this study, we described a new method used to release and collect the N -glycans from β -conglycinin, and the N -glycans existed in linear epitopes of β -conglycinin were identified. Glycopeptides hydrolyzed from β -conglycinin were purified by cotton hydrophilic chromatography. Trifluoromethylsulfonic acid was then used to release glycans from glycopeptides, and new glycopeptides containing one single N -acetyl- D -glucosamine (GlcNAc) moiety were then utilized for mass spectrometry. Five glycosylation sites (Asn-199, Asn-455, Asn-215, Asn-489 and Asn-326) and 22 kinds of glycopeptides were identified. It is noteworthy that the peptide VVN # ATSNL (where # represents for the glycosylation site) was analyzed to be both glycopeptide and linear epitope. Our results provided a new method for the N -glycoform analysis of food allergens, and laid a foundation for understanding the relationship between glycosylation and food allergy.