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Table 4 Binding affinity of IgG-Fc (upper rows) or IgG-Fab (lower rows) for PAB01 (WT) and PAB02 (D36H)

From: Engineered protein A ligands, derived from a histidine-scanning library, facilitate the affinity purification of IgG under mild acidic conditions

Protein

pH = 7.4

pH = 5.0

KD(×10−9) (M−1)

KDratio

KD(×10−9) (M−1)

KDratio

PAB01 (WT)

12.8

-

1080

-

PAB02 (D36H)

15.5

1.2

1350

1.3

 

pH = 7.4

pH = 5.0

Protein

K D (×10 −5 ) (M −1 )

K D ratio

K D (×10 −5 ) (M −1 )

K D ratio

PAB01 (WT)

2.9

-

ND

-

PAB02 (D36H)

6.7

2.3

ND

-

  1. The binding assay was performed under neutral (pH = 7.4) and acidic (pH = 5.0) conditions using SPR. IgG-Fc or IgG-Fab was immobilized on the surface of a CM5 sensor chip (GE Healthcare).
  2. KD, equilibrium dissociation constant: KD ratio, (KD of histidine-substituted PAB variant)/(KD of Wild-type PAB or PAZ): ND, not determined.
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Journal of Biological Engineering

ISSN: 1754-1611