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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

  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)
30.5 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
15 mg/mL-R 0.98 78.4 61.5 144 1.5 96.7 1.7
Polishing FT mode:
Capto adhere,
pH 6.8
10 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)
30.5 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
35 mg/mL-R 1.51 90.6 71.0 164 2.3 96.9 0.8
Polishing FT mode:
Capto adhere,
pH 6.8
30 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)
30.5 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
60 mg/mL-R 1.67 90.4 70.9 345 2.4 96.9 0.7
Polishing FT mode:
Capto adhere,
pH 6.8
45 mg/mL-R 0.65 79.4 56.3 88 0.3 98.6 1.1