Table 1.
Overview of used methods
| Nr. | Method | Lab | Description | Sample[µg] |
|---|---|---|---|---|
| 1 | HILIC(2-AB) | 1 | 2-AB labeling of released glycans; separation with HILIC-UPLC (LIF detection) | 200 |
| 2 | ESI-MS Heavy Chain | 1 | Reduction of mAb with TCEP and direct infusion with ESI-MS | 250 |
| 3 | ESI-MS after IdeS | 1 | Digestion with IdeS-Protease (Fabricator®) and direct infusion with ESI-MS | 100 |
| 4 | ESI-MS Glycopeptides | 1 | Tryptic digestion and direct infusion with ESI-MS after purification with Sepharose beads (HILIC) | 50 |
| 5 | LCMS with Orbitrap | 1 | LCMS after tryptic digestion with an Orbitrap™ mass spectrometer (Thermo Fisher™ Scientific Inc.) | 350 |
| 6 | LCMS with Q-TOF | 1 | LCMS after tryptic digestion with mass spectrometer Synapt® G2 q-TOF (Waters Corp.) | 350 |
| 7 | Nano-LCMS with Q-TOF | 2 | LCMS of tryptic digestion with mass spectrometer Maxis impact™ q-TOF, (Bruker Corp.) | 20 |
| 8 | PGC-MS | 1 | Release of glycans with PNGase F, separation with PGC coupled to ESI-MS | 450 |
| 9 | Positive MALDI-MS Glycopeptides | 2 | Tryptic digestion, HILIC-SPE and positive ion mode MALDI-TOF-MS | 20 |
| 10 | Negative MALDI-MS Glycopeptides | 2 | Tryptic digestion, HILIC-SPE and negative ion mode MALDI-TOF-MS | 20 |
| 11 | MALDI-MS Glycans | 1 | Release of glycans with PNGase F and measurement with MALDI-TOF-MS | 75 |
| 12 | MALDI-MS Stabilized Glycans | 2 | Release of glycans with PNGase F and measurement with MALDI-TOF-MS after ethyl esterification | 50 |