Table 1.
Binding kinetics of anti-hTNF-α variable new antigen receptor using Octet® RED96 system.
| Binding domain | Kon (M−1s−1) | Koff (s−1) | KD (nM) | KD (nM) BIAcore |
|---|---|---|---|---|
| D1 | 3.6 × 105 | 1.7 × 10−2 | n/a | 47.5 |
| C4 | 1.1 × 106 | 8.0 × 10−2 | n/a | 73.5 |
| TNF30 | 8.2 × 104 | 1.4 × 10−3 | n/a | 16.7 |
| D1–D1 | 5.0 × 105 | 3.2 × 10−4 | 0.6 | n/a |
| D1–C4 | 1.8 × 105 | 1.1 × 10−4 | 0.6 | 5.0 |
| D1–BA11–D1 | 1.9 × 106 | 2.0 × 10−4 | 0.1 | 4.0 |
| D1–BA11–C4 | 1.8 × 105 | 4.8 × 10−5 | 0.3 | 0.6 |
For all measurements, streptavidin Dip and Read™ biosensors were used to immobilize biotinylated hTNF-α. All assessed domains were loaded at a top concentration of 100 nM followed by a twofold dilution series. Values obtained from BIAcore binding analysis are shown in italics, and constructs not tested are shown as “not available” (n/a).