Table 1. Purification of expressed Hpn.
The protein content was determined using a BCA protein concentration assay (Bio-Rad). Hpn was quantified from an immunoblot using the His·Tag® monoclonal antibody, normalized against a standard curve of different Hpn concentrations. The percentage Hpn in the fraction was calculated as follows: (mg of Hpn/mg of total protein)×100.
| Fraction | Total protein (mg) | Hpn (mg) | Hpn in fraction (%) | Fold purification |
|---|---|---|---|---|
| Crude extract | 5300 | 2.6 | 0.05 | 1 |
| Crude supernatant | 3279 | 2.5 | 0.08 | 1.6 |
| Ni2+ affinity | 2.7 | 2.3 | 85 | 1700 |
| Gel filtration | 1.5 | 1.4 | >97* | 1940 |
* As determined from a Coomassie Blue-stained gel.