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Table 1 Preliminary run for MabSelect PrismA, where 9–10 mg of bsAb monomer containing CCS was loaded per mL of resin, with 3 CVs of 50 mM Na-citrate pH 6.0 wash step followed by their respective elution conditions

From: Excellent removal of knob-into-hole bispecific antibody byproducts and impurities in a single-capture chromatography

  

Elution conditions

Monomer concentration (mg/mL)

Monomer recovery (%)

Purity (%)

HMW

Mono

LMW

FabscFv-KiH

CCS

0.70

34.9

28.9

36.2

MabSelect PrismA

eluate

pH gradient elution from pH 6.0 to pH 3.0 in 25 CVs

1.02

101.1

8.5

90.9

0.6

2-step elution:

     

pH 3.8 mock pool

0.21

44.4

1.7

97.6

0.8

pH 3.6 mock pool

0.50

50.1

6.0

94.0

0.1

2-step elution:

     

pH 3.6 mock pool

0.78

94.6

6.7

92.3

0.9

pH 3.4 mock pool

0.02

0.8

61.8

26.7

11.5

Fab2scFv-KiH

CCS

0.78

34.3

30.4

35.3

MabSelect PrismA

eluate

pH gradient elution from pH 6.0 to pH 3.0 in 25 CVs

0.98

92.9

10.9

87.2

1.8

2-step elution:

     

pH 3.8 mock pool

pH 3.6 mock pool

0.24

51.2

5.1

89.8

5.1

0.52

43.9

6.8

91.1

2.1

2-step elution:

     

pH 3.6 mock pool

1.11

93.5

7.7

89.4

2.9

pH 3.4 mock pool

0.02

0.6

47.4

24.4

28.2