Table 1. Relative distribution of xylosylated glycans from an Fc-fusion protein after removal of Fc N-linked glycans using PNGase F (panel A) and from the linker tryptic peptide (panel B).
| Table 1A | ||
|---|---|---|
| Mass (Da) | Glycoforms on the Reduced Protein Chain | Relative Distribution (%) |
| 37877.2 | Deglycosylated* | ~72 |
| 38009.2 | Deglycosylated + 132.1 Da | ~18 |
| 38171.2 | Deglycosylated + 294.1 Da | ~4 |
| 38464.1 | Deglycosylated + 585.5 Da | ~4 |
| 38590.6 | Deglycosylated + 712 Da | ~2 |
| Table 1B | ||
|---|---|---|
| Mass (Da) | Glycan Structure(s) on the Linker Peptide** | Relative Distribution (%) |
| 2147.97 | none | 36.5 |
| 2280.01 | Xyl | 19.9 |
| 2412.05 | Xyl and Xyl | 1 |
| 2442.07 | Xyl-Gal | 2.5 |
| 2604.12 | Xyl-Gal-Gal | < 1 |
| 2684.08 | Xyl-[PO3]-Gal-Gal | 1.2 |
| 2733.16 | Xyl-Gal-Neu5Ac | 19.3 |
| 2780.15 | Xyl-Gal-Gal-GlcA | 2.5 |
| 2813.13 | Xyl-[PO3]-Gal-Neu5Ac | 5.9 |
| 2860.11 | Xyl-[PO3]-Gal-Gal-GlcA | 3.3 |
| 2865.20 | Xyl and Xyl-Gal-Neu5Ac | 1.7 |
| 2945.17 | Xyl and Xyl-[PO3]-Gal-Neu5Ac | < 1 |
| 2992.16 | Xyl and Xyl-[PO3]-Gal-Gal-GlcA | < 1 |
| 3027.26 | Xyl-Gal and Xyl-Gal-Neu5Ac | < 1 |
| 3063.19 | Xyl-[PO3]-Gal-Gal-GlcA-GlcNAc | 1.5 |
| 3143.16 | Xyl-[PO3]-Gal-Gal-GlcA-GlcNAc+(SO3) | < 1 |
| 3160.19 | Xyl-Gal-Gal-GlcA-GlcNAc-GlcA | < 1 |
| 3195.22 | Xyl and Xyl-Gal-Gal-GlcA-GlcNAc+(SO3) | < 1 |
| 3275.19 | Xyl and Xyl-[PO3]-Gal-Gal-GlcA-GlcNAc+(SO3) | < 1 |
| 3318.34 | Xyl-Gal-Neu5Ac and Xyl-Gal-Neu5Ac | < 1 |
| 3365.34 | Xyl-Gal-Neu5Ac and Xyl-Gal-Gal-GlcA | < 1 |
| 3444.31 | Xyl-Gal-Neu5Ac and Xyl-[PO3]-Gal-Gal-GlcA | < 1 |
*
Followed by treatment with PNGase F; **Attached to Ser(s) in the linker peptide.