Table 1.
Glutamine controls recHCG glycoforms in shake flask cultures.
| Peak No. |
Glycan Structure |
Day 1 0 mM Gln |
Day 3 0 mM Gln |
Day 5 0 mM Gln |
Day 1 4 mM Gln |
Day 3 4 mM Gln |
Day 5 4 mM Gln |
Day 1 8 mM Gln |
Day 3 8 mM Gln |
Day 5 8 mM Gln |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | A1 | 15.8 ± 7.4 | 18.8 ± 3.4 | 11.5 ± 0.8 | 7.8 ± 7.6 | 11.5 ± 3.2 | 10.0 ± 1.0 | 6.5 ± 5.8 | 12.3 ± 3.3 | 10.8 ± 0.1 |
| 2 | A2 | 0.4 ± 0.0 | 0.7 ± 0.0 | 0.6 ± 0.1 | 0.2 ± 0.0 | 0.4 ± 0.0 | 0.9 ± 0.1 | 0.3 ± 0.0 | 0.5 ± 0.0 | 0.8 ± 0.0 |
| 3 | A1G1 | 2.9 ± 0.4 | 3.7 ± 0.4 | 4.2 ± 0.2 | 0.9 ± 0.4 | 1.2 ± 0.4 | 1.6 ± 0.0 | 1.1 ± 0.2 | 1.5 ± 0.1 | 1.7 ± 0.1 |
| 4 | M5 | 8.0 ± 0.5 | 8.0 ± 0.5 | 8.3 ± 0.4 | 1.8 ± 0.1 | 2.8 ± 0.0 | 3.8 ± 0.0 | 1.9 ± 0.0 | 3.3 ± 0.1 | 4.1 ± 0.1 |
| 5 | A2G1, A1G1S1 |
2.5 ± 0.5 | 3.0 ± 0.0 | 3.2 ± 0.3 | 2.4 ± 0.3 | 2.6 ± 0.3 | 3.6 ± 0.3 | 2.3 ± 0.5 | 3.0 ± 0.1 | 3.8 ± 0.0 |
| 6 | A2G1S1 | 2.0 ± 0.6 | 1.8 ± 0.3 | 2.8 ± 0.1 | 0.9 ± 0.1 | 1.5 ± 0.0 | 1.5 ± 0.1 | 1.0 ± 0.2 | 1.3 ± 0.1 | 1.4 ± 0.1 |
| 7 | FA2G1 | 0.8 ± 0.5 | 1.0 ± 0.0 | 0.9 ± 0.2 | 1.2 ± 0.3 | 1.1 ± 0.1 | 2.4 ± 0.1 | 1.2 ± 0.2 | 1.2 ± 0.1 | 2.5 ± 0.0 |
| 8 | A2G2, FA2G1S1 |
13.5 ± 1.3 | 12.6 ± 1.2 | 16.4 ± 0.1 | 7.3 ± 0.4 | 9.3 ± 0.0 | 8.3 ± 0.5 | 7.8 ± 0.2 | 8.5 ± 0.2 | 7.9 ± 0.4 |
| 9 | FA2G2, A2G2S1 |
20.7 ± 2.0 | 19.6 ± 1.5 | 22.6 ± 0.7 | 22.5 ± 0.5 | 23.7 ± 0.1 | 21.3 ± 0.4 | 23.9 ± 0.1 | 22.6 ± 0.8 | 20.3 ± 0.6 |
| 10 | A2G2S2, FA2G2S1 | 19.5 ± 1.2 | 17.1 ± 1.1 | 17.6 ± 0.4 | 32.6 ± 5.3 | 26.1 ± 0.9 | 26.3 ± 0.2 | 32.7 ± 4.6 | 27.1 ± 1.3 | 25.6 ± 0.4 |
| 11 | A3G2S1, FA2G2S2 | 8.8 ± 0.2 | 9.7 ± 0.7 | 7.6 ± 0.3 | 14.6 ± 2.3 | 12.4 ± 1.6 | 13.2 ± 1.6 | 14.1 ± 0.3 | 12.3 ± 1.1 | 13.2 ± 0.9 |
| 12 | A3G2S2, A3G3S1 |
1.9 ± 0.5 | 1.5 ± 0.1 | 1.7 ± 0.1 | 2.5 ± 0.2 | 2.4 ± 0.2 | 2.2 ± 0.3 | 2.4 ± 0.3 | 2.0 ± 0.1 | 2.5 ± 0.1 |
| 13 | FA3G3S1, A3G3S2 | 1.3 ± 0.2 | 1.0 ± 0.1 | 1.3 ± 0.1 | 2.1 ± 0.1 | 2.1 ± 0.2 | 1.9 ± 0.1 | 1.9 ± 0.1 | 1.8 ± 0.0 | 2.3 ± 0.0 |
| 14 | A3G3S3, FA3G3S2 | 1.1 ± 0.0 | 0.9 ± 0.1 | 1.1 ± 0.0 | 2.3 ± 0.4 | 1.9 ± 0.2 | 2.0 ± 0.1 | 2.2 ± 0.3 | 1.8 ± 0.1 | 2.0 ± 0.0 |
| 15 | FA3G3S3 | 0.6 ± 0.0 | 0.5 ± 0.2 | 0.4 ± 0.1 | 0.8 ± 0.0 | 0.9 ± 0.2 | 1.0 ± 0.2 | 0.6 ± 0.2 | 0.8 ± 0.2 | 1.1 ± 0.0 |
The relative structures and proportions of each peak of the HCG N-linked glycan NP-HPLC pool and on day 1, 3 and 5 of batch culture with varying concentrations of glutamine. Numbers in bold indicates significant difference, p < 0.05, n = 6.