TABLE 1.
Characterization of Evasin-4-Fc mutants
Inhibitory properties were determined by inhibition of L1.2/CCR5 (40 nm CCL3 or 40 nm CCL5) or L1.2/CCR2 (10 nm CCL8) chemotaxis and are characterized by the associated IC50. Dissociation constants (KD) were measured by SPR. −, no inhibition of chemotaxis or no binding; n.d., not determined as the fitting was not good enough to determine an accurate KD due to low signal and chemokine oligomerization.
| ka × 103 | kd × 10−4 | KD | IC50 | |
|---|---|---|---|---|
| m−1 s−1 | s−1 | nm | nm | |
| CCL3 | ||||
| Evasin-4-Fc | 5 | 4 | 71 | 11 |
| E16A | 4 | 4 | 81 | 17 |
| Y19A | 5 | 15 | 283 | 37 |
| E16A/Y19A | − | − | − | 36 |
| Y28Q/N60D | 29 | 89 | 310 | 133 |
| CCL5 | ||||
| Evasin-4-Fc | 31 | 84 | 274 | 97 |
| E16A | 36 | 134 | 377 | 176 |
| Y19A | n.d | n.d. | n.d. | 7598 |
| E16A/Y19A | − | − | − | − |
| Y28Q/N60D | 34 | 248 | 726 | 380 |
| CCL8 | ||||
| Evasin-4-Fc | 17 | 8 | 47 | 0.8 |
| E16A | 13 | 15 | 112 | 1.1 |
| Y19A | 26 | 55 | 209 | 1.7 |
| E16A/Y19A | − | − | − | 17.3 |
| Y28Q/N60D | 31 | 78 | 248 | 2.0 |