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Table 7 5-mL validation runs of the fully optimised process for both FabscFv-KiH and Fab2scFv-KiH

From: Effective flow-through polishing strategies for knob-into-hole bispecific antibodies

   Load Monomer concentration (mg/mL) Step monomer recovery (%) Overall monomer recovery (%) Leached Protein A (ppm) HCP (ppm) hcDNA (ppm) Purity (%)
HMW Mono LMW
  CCS 0.56 - - N.D 1,307,373 67,006.3 30.8 34.0 35.2
FabscFv-KiH Post-Protein A eluate
(pH 4.7 wash)
31.5 mg/mL-R 3.68 91.1 91.1 5 2350 0.5 4.8 94.7 0.5
Polishing FT mode:
Capto Butyl ImpRes,
pH 4.0
40 mg/mL-R 2.25 92.7 84.4 N.D 189 0.5 1.8 98.1 0.1
Polishing FT mode:
Capto adhere,
pH 6.8
30 mg/mL-R 0.76 88.7 74.9 N.D 87 0.2 0.6 99.3 0.1
  CCS 0.72 - - N.D 1,077,136 54,133.9 33.5 31.1 35.4
Fab2scFv-KiH Post-Protein A eluate
(pH 4.1 wash)
30.5 mg/mL-R 1.75 78.4 78.4 6 2064 0.5 5.1 93.2 1.7
Polishing FT mode:
Capto Butyl ImpRes,
pH 4.0
60 mg/mL-R 1.52 93.0 72.9 N.D 153 0.3 2.6 96.6 0.8
Polishing FT mode:
Capto adhere,
pH 6.8
45 mg/mL-R 0.73 82.1 59.9 N.D 59 0.1 0.3 99.2 0.5
  1. Representative post-Protein A eluates are reported in the table, as multiple post-Protein A eluates from 5-mL column are pooled together to generate sufficient materials for the subsequent polishing steps