Table 2. Purification of recombinant cABC I.
| Fraction | Protein yield (mg) | Specific activity (1 mg/ml C6S) (units/mg) | x-Fold purification* |
|---|---|---|---|
| Crude lysate | 195 | 92.1 | |
| Elution from Ni2+ column | 35 | 197.9 | 2.1 |
| Thrombin cleavage† | 35 | 230.6 | 2.5 |
* The x-fold purification was determined relative to the specific activity measured for crude lysate.
† Cleavage of the His6 tag from the recombinant protein was confirmed by Western blot analysis using an anti-(His tag) antibody (Amersham Biosciences). No detectable difference in activity against GalAG substrates was observed between the recombinant cABC I and its His-tag-cleaved counterpart.