DE52 diethylaminoethyl cellulose (DEAE-C) anion-exchange column chromatography was performed using HKBML, TK, RAJI, OYB, HBL1, and A4/FUK (Additional file
3: Figure S2A and Additional file
4: Figure S3A), and MTX-resistant HKBML, TK, and RAJI (Additional file
3: Figure S2A). Coupled with standard unit for sialylated sugar chains, the peaks representing non-sialylated (S0) to 4-sialylated (S4) sugar chains were detected (Additional file
3: Figure S2A and Additional file
4: Figure S3A). The left-side peaks at the S1 peak indicated the
N-oligosaccharides coupling with an unknown acidic group, and the left-side peaks at the S2 peak indicated the
N-mono-sulfated oligosaccharides (Additional file
2: Figure S1A and Additional file
3: Figure S2A). Fractions eluting at wide-range retention times of S0 were further analyzed using normal-phase HPLC in neutral sugar chains and sialylated sugar chains (Additional file
3: Figures S2B–G and Additional file
4: Figure S3B, C). Fractions eluting at retention times of M4-M9 were collected and analyzed, and the oligostructures included in the peaks were determined by MALDI/TOF-MS, then referred to the oligosaccharide structures in Additional file
5: Figure S4. High mannose type oligosaccharides, including M4B, M5A, M6B, M7A, M7B, M8A, and M9A were determined with mannose unit standard in all samples examined (Additional file
3: Figure S2B–G, Additional file
4: Figure S3B, C, and Additional file
5: Figure S4). Interestingly, biantennary bigalactosylated structure with α1,6-fucosylation, A2G2F, and the
N-acetylglucosamine-bisecting structure, A2G2FB as complex types, were also detected in neutral and sialylated oligosaccharides in cells derived from HKBML, TK, and RAJI (Additional file
3: Figure S2B–G and Additional file
5: Figure S4). Further, tetra-antennary tetra-galactosylated structure with α1,6-fucosylation, A4G4F, was only detected in cells from RAJI (Additional file
3: Figure S2F, G and Additional file
5: Figure S4). These complex types were not detected in HBL1, A4/FUK, and OYB (Additional file
4: Figure S3B, C).