Table 13.
Protein loss during purification of TcdB.
| Stage | Purification Step | Volume (mL) | TcdB | Starting Material (%) b | Purity c | |
|---|---|---|---|---|---|---|
| µg/mL | Total (mg) | |||||
| 1 | Supernatant | 810 a | 17.2 | 13.9 | 100 | N/A |
| 2 | Diafiltration | 230 a | 39.8 | 9.2 | 66.2 | 3.9% |
| 3 | Q Sepharose | 40 | 149.4 | 6.0 | 43.2 | 65.6% |
| 4 | Mono Q | 18.6 | 266.6 | 5.0 | 36.0 | 95.2% |
| 5 | Ultrafiltration | 0.26 | 19,425.9 | 5.0 | 36.0 | 98% |
| 6 | Superdex 200 | 3 | 1167.0 | 3.5 | 25.2 | >99% |
a Volume containing TcdA and TcdB. Subsequent volumes contain only TcdB. b Calculated as the total amount of TcdB at each stage relative to the filtered supernatant (stage 1). c Purity from SDS-PAGE (Supplementary Figure S2) was estimated using the lane profile function of Image Lab 6.1 software function on a Bio-Rad Gel Doc Imager.