Table 1.
Summary of the analytical methodologies and instruments used for the N- and O-glycan profiling of glycoproteins in lyophilized cell pellets
| Lab code |
N-glycan preparation |
N-glycan derivatization |
O-glycan preparation |
O-glycan derivatization |
Analysis strategy |
MS instrument |
|---|---|---|---|---|---|---|
| Lab A | Pr/Pn | OS/PA | Pr/Pn/Hy | OS/PA | HPLC: AE/RP-LC/FL, MALDI-TOF-MS (+ ion) | Shimadzu AXIMA-CFR MALDI-TOF |
| Lab B | Pn | OS/AA | Pr/AGC | OS/AA | HPLC: Se-LC/FL, MALDI-TOF-MS(MSn) (+ ion) | Shimadzu AXIMA Resonance MALDI-QIT-TOF |
| Lab C | RA/Pr/Pn | OS/PM | RA/Pr/Pn/ β-elim | OSa/PM | MALDI-TOF-MS (+ ion) | ABI Voyager DE Pro MALDI-TOF |
| Lab D | RA/Pr/Pn | OSa/PM | RA/Pr/Pn/β-elim | OSa/PM | MALDI-TOF-MS (+ ion) | Bruker Ultraflex I MALDI-TOF |
| Lab E | RA/Pr/Pn | OSa/PM | RA/Pr/Pn/β-elim | OSa/PM | MALDI-TOF-MS(MSn) (+ ion) | Bruker Reflex IV MALDI-TOF, Shimadzu AXIMA QIT MALDI-QIT-TOF |
| Lab F | Pn | OSa | Pn/β-elim | OSa | PGC-LC-ESI-MS(/MS) (− ion) | Agilent LC/MSD Trap XCT Plus Series 1100 |
| Lab G | RA/Pr/Pn | Osa | Not participated | Not participated | PGC-LC-ESI-MS (+/− ion) | Thermo Fisher Scientific LTQ FT |
AA 2-aminobenzoic acid, AE anion exchange, AGC AutoGlycoCutter, β-elim reductive β-elimination, PGC porous graphitic carbon column, FL fluorescence, Gp glycopeptide, Hy hydrazinolysis, OS oligosaccharides (reducing sugar), OSa oligosaccharide alditols, PA pyridylamination, PM permethylation, Pn peptide-N-glycosidase treatment, N-glycan removal, Pr proteolytic digestion, RA reduction and cysteine derivatization, RP reverse phase, Se serotonin chromatography, +/− ion positive/negative ion mode