Table 1.
Summary of caffeine sensitivity (EC50 values) for representative acVHH variants and biCOSMO variants tested in the study. n = 16 cells from three independent assays. Variants with functional validation were highlighted in bold
| Variantsa) | EC50 |
|---|---|
| WT | (567.5 ± 1.6) × 10−9 m |
| Y104W (COSMO) | (95.1 ± 1.2) × 10−9 m |
| Y104F | (3990.2 ± 4.7) × 10−9 m |
| Y34W | >20 × 10−6 m |
| Y34F | No binding |
| Y108W | >2 × 10−6 m |
| Y108F | No binding |
| Y61W | >1 × 10−6 m |
| Y61F | No binding |
| F39W | No binding |
| F39Y | No binding |
| W111F | No binding |
| W111Y | No binding |
| M63L | No binding |
| biCOSMO‐S | (16.9 ± 2.0) × 10−9 m |
| biCOSMO‐L | Intramolecular binding |
| 2 × WT acVHH‐S | (530.9 ± 1.6) × 10−9 m |