TABLE 3.
Kinetics and binding of TNFα mutants with infliximab Fab
Kinetics and binding of TNFα mutant and infliximab Fab were analyzed using a BIAcore T100. Wild-type TNFα and the mutants were passed over the immobilized infliximab Fab surface, and the data were globally analyzed using a simultaneous fit for both dissociation (kd) and association (ka). The value for KD was calculated as kd/ka. Typical error levels for the kd and ka values are less than ±15%.
| ka | kd | KD | |
|---|---|---|---|
| m−1·s−1 | s−1 | m | |
| WT TNFα | 3.50 ± 0.52 × 104 | 3.0 ± 0.45 × 10−4 | 8.70 ± 1.3 × 10−9 |
| TNFα(EF-4G) | 0.96 ± 0.14 × 104 | 1.7 ± 0.25 × 10−2 | 1.78 ± 0.27 × 10−6 |
| TNFα(EF-11SG) | 3.00 ± 0.45 × 104 | 5.0 ± 0.75 × 10−2 | 1.80 ± 0.27 × 10−6 |
| TNFα(Q67A) | 5.86 ± 0.88 × 103 | 1.80 ± 0.27 × 10−4 | 3.07 ± 0.46 × 10−8 |
| TNFα(K112A) | 1.67 ± 0.25 × 104 | 4.74 ± 0.71 × 10−4 | 2.84 ± 0.43 × 10−8 |
| TNFα(R138A) | 1.76 ± 0.26 × 102 | 1.69 ± 0.25 × 10−3 | 9.61 ± 1.44 × 10−6 |
| TNFα(Y141A) | 2.02 ± 0.30 × 102 | 1.07 ± 0.16 × 10−3 | 5.32 ± 0.80 × 10−6 |