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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