Table 4.
FIX purification
| Fraction name | FIX:Ag, IU/ml | Fraction volume, ml | FIX:C, IU/ml | FIX:Ag, IU |
Total protein quantified by UV spectrophotometric assay, mg |
FIX:C, IU | FIX:C/ protein, IU/mg | FIX:Ag/ /protein, IU/mg |
Yield of the stage with respect to FIX:Ag |
Overall yield with respect to FIX:Ag |
Percentage of FIX with propeptide, % |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Culture medium | 6.04 | 115 | 3.6 | 695 | - | 414 | - | - | - | - | 2,6 |
| Capto MMC flow-through and eluate fractions | 0.166 | 273 | - | 45 | - | - | - | - | 7% | - | - |
| Capto MMC eluate | 64.79 | 9 | - | 583 | 4.28 | - | - | 136 | 84% | 84% | - |
| Capto Q flow-through fraction | 0.07 | 37 | - | 3 | 0.69 | - | - | 4 | 0.4% | - | - |
| Capto Q eluate 200 mM NaCl | 0.05 | 12.5 | - | 1 | 0.48 | - | - | - | - | - | |
| Capto Q eluate 10 mM CaCl2 | 31.75 | 9.2 | 25.2 | 292 | 1.51 | 232 | 154 | 194 | 50% | 62% | 2 |
| Capto Q, 150 mM NaCl + 10 mM CaCl2 | 12.95 | 10.5 | - | 136 | 0.88 | - | - | 154 | 23% | ||
| Capto Q 200 mM NaCl + 10 mM CaCl2 | 4.67 | 10.7 | <1 | 50 | 0.83 | <10.7 | <12 | 60 | 9% | - | 5.8 |
| Capto Q 500 mM NaCl + 10 mM CaCl2 | 13.87 | 4 | - | 55 | 0.57 | - | - | 98 | 10% | - | - |
| Capto Heparin flow-through fraction | 0.72 | 18 | - | 13 | - | - | - | - | 3% | - | - |
| Capto Heparin eluate | 36.10 | 9 | 38.6 | 325 | 1.46 | 347 | 237 | 222 | 76% | 32% | <2 |