Table 3.
Purification of aFGF from 6HFh8-aFGF fusion protein expressed in E. coli
| Purification step | Concentration (mg/ml)a | Volume (ml) | Total protein (mg) | Fusion protein (and tag-free) (mg) | Purity (%)a | Yield (%)b |
|---|---|---|---|---|---|---|
| Crude extract | 4.6 ± 0.1 | 70.0 | 322.7 ± 8.9 | 153.0 ± 2.8 (81.0 ± 1.5) | 47.4 | 100.0 |
| HisTrap, 5 ml | 2.5 ± 0.0 | 67.3 | 165.6 ± 2.9 | 142.5 ± 2.5 (75.5 ± 1.3) | 86.1 | 93.2 |
| HisTrap, 5 ml | 1.6 ± 0.0 | 50.1 | 77.9 ± 2.5 | NA (72.3 ± 2.3) | 92.8 | 89.2 |
| HiTrap CM, 5 ml | 0.7 ± 0.0 | 80.5 | 59.6 ± 2.3 | NA (59.6 ± 2.3) | 100.0 | 73.5 |
The data are shown as mean ± standard deviation from triplicate independent experiments
aPurity was analyzed by densitometry using ImageJ and C18 RP HPLC
bYield was calculated by dividing the tag-free protein of each purification product by tag-free protein in crude extract