Table 3.
Comparison of Tm, refolding and binding kinetics for EBOV GP binders and their derivatives
| Clone name | Tm (°C) | Refolding (%) | ka (1/Ms) | kd (1/s) | KD (M) |
|---|---|---|---|---|---|
| EBOV-GP-G6 | 72 | 77 | 7.0 E+04 | 1.5E−04 | 2.0E−09 |
| EBOV-GP-G6-neg+ | 87 | 67 | 3.5E+03 | 1.5E−04 | 4.4E−08 |
| EBOV-GP-H7 | 70 | 97 | 1.7E+04 | 3.0E−04 | 1.7E−08 |
| EBOV-GP-H7-neg+ | 87 | 85 | 3.5E+04 | 1.4E−03 | 4.0E−08 |
| EBOV-GP-A8 | 57 | 0 | 3.1E+04 | 7.2E−05 | 2.3E−09 |
| EBOV-GP-A8-neg+ | 73 | 0 | 6.1E+04 | 5.8E−05 | 9.5E−10 |
| EBOV-GP-A8-fneg+ | 74 | 66 | 1.3E+05 | 9.1E−05 | 6.9E−10 |
Three of the GP binding sdAbs showing the highest melting temperatures and/or best affinities were subject to mutagenesis to improve their stability. Melting temperature and refolding ability were measured by CD