Table 6 Evaluation of different load amounts on polishing steps for Fab2scFv-KiH
From: Effective flow-through polishing strategies for knob-into-hole bispecific antibodies
| Â | Fab2scFv-KiH | Load | Monomer concentration (mg/mL) | Step monomer recovery (%) | Overall monomer recovery (%) | HCP (ppm) | Purity (%) | ||
|---|---|---|---|---|---|---|---|---|---|
HMW | Mono | LMW | |||||||
|  | CCS | - | 0.72 | – | – | 1,329,885 | 33.5 | 31.1 | 35.4 |
Process 1 | Post-Protein A eluate (pH 4.1 wash) | 30.5Â mg/mL-R | 2.92 | 78.4 | 78.4 | 1526 | 6.5 | 92.1 | 1.4 |
Polishing FT mode: Capto Butyl ImpRes, pH 4.0 | 15Â mg/mL-R | 0.98 | 78.4 | 61.5 | 144 | 1.5 | 96.7 | 1.7 | |
Polishing FT mode: Capto adhere, pH 6.8 | 10Â mg/mL-R | 0.16 | 56.0 | 34.4 | 57 | 0.0 | 97.8 | 2.1 | |
Process 2 | Post-Protein A eluate (pH 4.1 wash) | 30.5Â mg/mL-R | 2.92 | 78.4 | 78.4 | 1526 | 6.5 | 92.1 | 1.4 |
Polishing FT mode: Capto Butyl ImpRes, pH 4.0 | 35Â mg/mL-R | 1.51 | 90.6 | 71.0 | 164 | 2.3 | 96.9 | 0.8 | |
Polishing FT mode: Capto adhere, pH 6.8 | 30Â mg/mL-R | 0.49 | 75.1 | 53.3 | 84 | 0.2 | 98.9 | 0.9 | |
Process 3 | Post-Protein A eluate (pH 4.1 wash) | 30.5Â mg/mL-R | 2.92 | 78.4 | 78.4 | 1526 | 6.5 | 92.1 | 1.4 |
Polishing FT mode: Capto Butyl ImpRes, pH 4.0 | 60Â mg/mL-R | 1.67 | 90.4 | 70.9 | 345 | 2.4 | 96.9 | 0.7 | |
Polishing FT mode: Capto adhere, pH 6.8 | 45Â mg/mL-R | 0.65 | 79.4 | 56.3 | 88 | 0.3 | 98.6 | 1.1 | |