Table 2.
Quantitative evaluation of method performance. Each analytical method was applied in 2 separate series 6 replicates per batch. Relative abundance of the various glycan species are given in percent, with standard deviations in parentheses. Key: H, hexose; N, N-acetylhexosamine; F, deoxyhexose; S, N-acetylneuraminic acid (sialic acid); G0F-N, agalacosylated, core-fucosylated, monoantennary species, etc.; n.d.: not detected; n.a.: not applicable. n,q.: not quantifiable
| Glycan species | Structural scheme | HILIC(2-AB)Reference Method | ESI-MS Heavy Chain | ESI-MS after IdeS | ESI-MS Glycopeptides | LCMS with Orbitrap | LCMS with Q-TOF | nano-LCMS with Q-TOF | PGC-MS | Positive MALDI_MS Glycopeptides | Negative MALDI-MS Glycopeptides | MALDI-MS Glycans | MALDI-MS Stabilized Glycans |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| G0F[H3N4F1] | 35.5 (0.1)35.3 (0.1) | 34.9 (0.2)34.7 (0.2) | 31.3 (0.5)32.0 (0.3) | 29.3 (0.2)30.6 (0.7) | 29.8 (0.2)32.4 (0.3) | 29.7 (0.4)29.9 (0.3) | 32.4 (0.5)33.6 (0.9) | 36.5 (0.4)36.2 (0.2) | 37.8 (0.9)38.2 (0.3) | 38.0 (0.9)38.4 (0.1) | 37.3 (0.2)37.6 (0.5) | 30.6 (0.7)31.2 (0.2) | |
| G1F[H4N4F1] | 43.4 (0.1)43.3 (0.1) | 45.5 (0.2)45.6 (0.2) | 39.5 (0.3)39.6 (0.3) | 40.6 (1.0)38.6 (0.4) | 38.4 (0.1)39.2 (0.3) | 40.1 (0.4)40.1 (0.3) | 42.4 (0.3)40.5 (1.8) | 42.7 (0.4)42.6 (0.2) | 46.6 (0.5)46.5 (0.1) | 47.0 (0.5)46.9 (0.2) | 44.4 (0.6)44.4 (0.6) | 46.1 (0.7)47.9 (0.3) | |
| G2F[H5N4F1] | 9.5 (<0.1)9.6 (0.1) | 10.7 (0.1)11.1 (0.1) | 11.9 (0.5)11.1 (0.3) | 9.8 (0.2)8.6 (0.4) | 10.1 (0.1)9.3 (0.1) | 9.0 (0.1)9.0 (0.1) | 9.7 (0.3)9.0 (0.3) | 9.2 (0.1)9.2 (0.1) | 7.3 (0.5)7.3 (0.2) | 7.0 (0.4)7.0 (0.1) | 7.0 (0.1)6.6 (0.3) | 9.9 (0.5)9.0 (0.3) | |
| G0[H3N4] | 4.6 (0.1)4.7 (0.1) | 9.0 (0.3)8.5 (0.2) | 6.0 (0.1)6.0 (0.1) | 5.1 (0.3)5.9 (0.3) | 6.1 (<0.1)6.6 (<0 .1) | 4.4 (0.1)4.6 (0.1) | 4.9 (0.3)5.8 (0.5) | 3.6 (0.1)3.8 (0.1) | 3.6 (0.2)3.8 (<0.1) | 3.4 (0.1)3.7 (0.1) | 5.0 (0.1)4.8 (0.2) | 4.3 (0.2)4.0 (0.2) | |
| G1[H4N4] | 3.3 (<0.1)3.4 (<0 .1) | n.d.n.d. | n.q.n.q. | 3.7 (0.2)3.8 (0.1) | 4.7 (0.1)4.8 (<0.1) | 3.0 (0.1)3.0 (0.2) | 3.3 (0.2)3.6 (0.2) | 2.5 (<0.1)2.6 (<0.1) | 2.1 (0.1)2.0 (0.1) | 2.0 (0.1)1.9 (<0.1) | 3.0 (0.1)3.0 (0.1) | 4.1 (0.1)3.8 (0.1) | |
| G2[H5N4] | 0.3 (<0 .1)0.4 (<0.1) | n.d.n.d. | n.d.n.d. | 1.1 (0.1)1.0 (0.1) | 0.7 (<0 .1)0.6 (0.1) | 0.4 (<0 .1)0.4 (<0.1) | 0.9 (0.1)0.9 (0.1) | 0.5 (<0 .1)0.4 (<0.1) | 0.3 (<0 .1)0.2 (<0.1) | 0.3 (<0 .1)0.2 (<0.1) | 0.5 (0.1)0.5 (0.1) | 1.5 (0.1)1.3 (0.2) | |
| G0F-N[H3N3F1] | 0.5 (<0.1)0.5 (<0.1) | n.d.n.d. | 1.1 (0.1)1.2 (0.1) | 2.7 (0.2)3.7 (0.2) | 2.5 (0.1)1.2 (<0.1) | 5.6 (0.1)5.6 (0.1) | 1.0 (<0.1)0.9 (<0.1) | 0.5 (<0.1)0.5 (<0.1) | 0.4 (<0.1)0.3 (<0.1) | 0.4 (<0.1)0.3 (<0.1) | 0.6 (0.1)0.8 (0.1) | <0.1 (<0.1)<0.1 (<0.1) | |
| G1F-N[H4N3F1] | n.d.(n.d.) | n.d.n.d. | 2.4 (0.1)2.6 (0.1) | 2.7 (0.2)2.4 (0.1) | 1.3 (<0.1)0.9 (<0.1) | 2.7 (0.1)2.6 (0.1) | 0.7 (<0.1)0.6 (<0.1) | 0.6 (<0.1)0.6 (<0.1) | 0.3 (<0.1)0.2 (<0.1) | 0.3 (<0.1)0.2 (<0.1) | 0.5 (<0.1)0.4 (<0.1) | 0.5 (<0.1)0.4 (0.1) | |
| G0-N[H3N3] | 0.4(<0.1)0.4(<0.1) | n.d.n.d. | 1.1 (0.1)1.1 (0.1) | 1.1 (0.1)1.3 (0.1) | 1.2 (<0.1)0.9 (<0.1) | 1.3 (0.1)0.9 (0.1) | 0.7 (<0 .1)0.7 (0.1) | 0.2 (<0.1)0.2 (<0.1) | 0.2 (<0.1)0.2 (<0.1) | 0.2 (<0.1)0.2 (<0.1) | 0.8 (0.1)1.0 (<0.1) | 0.2 (0.1)0.1 (<0.1) | |
| M5[H5N2] | 1.5 (<0.1)1.6 (<0.1) | n.d.n.d. | 2.4 (0.1)2.3 (0.1) | 1.9 (0.2)1.9 (<0.1) | 2.9 (0.1)3.1 (0.1) | 1.2 (0.1)1.3 (0.1) | 1.9 (<0.1)1.9 (0.1) | 0.5 (<0.1)0.5 (<0.1) | 1.1 (0.1)1.0 (<0.1) | 1.1 (0.1)1.0 (<0.1) | 0.9 (<0.1)0.9 (0.1) | 1.0 (0.1)0.9 (0.2) | |
| M6[H6N2] | 0.1 (<0.1)0.1 (<0.1) | n.d.n.d. | n.d.n.d. | 0.4 (0.1)0.3 (0.1) | 0.2 (<0.1)0.2 (<0.1) | 0.1 (<0.1)0.1 (<0.1) | n.d.n.d. | n.d.n.d. | n.q.n.q. | n.q.n.q. | n.d.n.d. | n.d.n.d. | |
| G1FS[H4N4FS1] | 0.2 (<0.1)0.2 (<0.1) | n.d.n.d. | 1.3 (0.1)1.4 (0.1) | 0.4 (0.1)0.4 (<0 .1) | 0.4 (<0.1)0.1 (<0.1) | 0.4 (<0.1)0.3 (<0.1) | n.d.n.d. | 0.9 (<0.1)0.9 (<0.1) | n.q.n.q. | n.q.n.q. | n.d.n.d. | 0.3 (0.1)0.2 (<0.1) | |
| G2S1F[H5N4FS1] | 0.7 (<0.1)0.7 (0.1) | n.d.n.d. | 1.9 (0.2)1.8 (0.1) | 0.6 (0.1)0.6 (0.1) | 1.1 (<0 .1)0.4 (<0.1) | 0.9 (0.1)0.6 (0.1) | 1.4 (0.1)1.5 (0.1) | 2.1 (0.1)2.2 (0.1) | n.q.n.q. | n.q.n.q. | n.d.n.d. | 0.7 (0.1)0.5 (<0.1) | |
| G2S2F[H5N4FS2] | 0.1 (<0.1)0.1(<0.1) | n.d.n.d. | 1.0 (0.1)1.0 (0.1) | 0.2 (0.1)0.2 (<0.1) | 0.2(<0.1)<0.1 (<0.1) | 0.3 (<0.1)0.3 (<0.1) | 0.4 (0.1)0.5 (0.1) | n.d.n.d. | n.d.n.d. | n.d.n.d. | n.d.n.d. | 0.3 (0.1)0.1 (0.1) |