Table 2.
Prokaryotic expression, refolding, and purification of recombinant mPRDC
| Purification step | Total Protein (mg) b | Total PRDC (mg) b | Overall yield (%) | Purity (%) c |
|---|---|---|---|---|
| Bacterial lysatea | 531.9 | 50 | 100 | 9.4 |
| Unwashed Inclusion bodies | 434.7 | 46.5 | 93 | 10.7 |
| Washed inclusion bodies | 76.2 | 44.1 | 88.2 | 57.9 |
| Sephacryl S-200 (IB) | 20.5 | 17.4 | 34.8 | 84.6 |
| Hiprep SP (IB) | 25.8 | 21.3 | 42.6 | 82.6 |
| Hiprep SP (IB/Sephacryl S-200) | 4.8 | 4.6 | 9.2 | 95.4 |
| Refold and C18 (Sephacryl S-200 only) | 3.4 | 3.1 | 6.2 | 89.6 |
| Refold and C18 (Hiprep SP only) | 4.9 | 4.1 | 8.2 | 82.5 |
| Refold and C18 (Sephacryl S-200+ Hiprep SP) | 1.3 | 1.26 | 2.6 | >97 |
a
Results are given for 8 g of wet weight cells derived from 4 L of E. coli culture.
b
Protein concentration from pooled samples determined by the BCA assay using bovine serum albumin as a standard protein.
c
The purity was determined by using SDS-PAGE densitometry software ImageJ (http://rsb.info.nih.gov/ij/).