Table 3 The purity profile, monomer concentration and recovery of each step of the Protein A run for both FabscFv-KiH and Fab2scFv-KiH in 1-mL columns with a load of 50% of QB10 at 6-min residence time, with the introduction of an intermediate low-pH wash step
Conditions | Monomer concentration (mg/mL) | Monomer recovery (%) | Purity (%) | |||||
|---|---|---|---|---|---|---|---|---|
HMW | Mono | LMW | ||||||
1-mL column | FabscFv-KiH | CCS | – | 0.69 | – | 30.8 | 35.5 | 33.8 |
MabSelect PrismA | FT | 0.02 | 2.4 | – | – | – | ||
pH 6.0 wash | 0.01 | 0.1 | – | – | – | |||
pH 4.7 wash | 0.03 | 1.2 | 7.6 | 48.2 | 44.2 | |||
pH 3.6 peak (UV280 > 50 mAu) | 3.65 | 90.6 | 6.7 | 92.9 | 0.4 | |||
pH 3.6 tail (UV280 < 50 mAu) | 0.04 | 1.2 | 29.7 | 67.7 | 2.6 | |||
Fab2scFv-KiH | CCS | - | 0.72 | - | 33.5 | 31.1 | 35.4 | |
MabSelect PrismA | FT | 0.02 | 2.7 | - | - | - | ||
pH 6.0 wash | 0.01 | 0.1 | - | - | - | |||
pH 4.3 wash | 0.02 | 0.7 | 6.1 | 18.3 | 75.6 | |||
pH 3.6 peak (UV280 > 50 mAu) | 2.04 | 86.6 | 5.8 | 92.3 | 1.9 | |||
pH 3.6 tail (UV280 < 50 mAu) | 0.06 | 1.1 | 16.8 | 78.9 | 4.3 | |||
MabSelect PrismA | FT | 0.01 | 1.9 | - | - | - | ||
pH 6.0 wash | 0.01 | 0.1 | - | - | - | |||
pH 4.1 wash | 0.15 | 4.9 | 17.8 | 62.3 | 19.9 | |||
pH 3.6 peak (UV280 > 50 mAu) | 1.56 | 78.7 | 6.4 | 92.2 | 1.4 | |||
pH 3.6 tail (UV280 < 50 mAu) | 0.07 | 1.1 | 12.2 | 87.1 | 0.7 | |||