Table 2.
Framework optimization of nanobodies generated against wildtype SARS-CoV-2 spike protein.
| Original Mutation | TM (°C) | KD (M) | ||
|---|---|---|---|---|
|
| ||||
| LaG21L69F | Wildtype | S1–8 | 74.0 | 3.8 |
| Mutant | S1–8L70F | 83.0 | 3.2 | |
|
| ||||
| Wildtype | S1–28 | 66.0 | 0.6 | |
| Mutant | S1–28L70F | 77.5 | 0.09 | |
|
| ||||
| Wildtype | S1–36 | 66.5 | 0.2 | |
| Mutant | S1–36L70F | 76.0 | undetectable | |
|
| ||||
| LaG16E91K | Wildtype | S1–40 | 57.0 | undetectable |
| Mutant | S1–40S89K | 59.5 | 0.5 | |
|
| ||||
| Wildtype | S1-RBD-14 | 65.0 | 25 | |
| Mutant | S1-RBD-14E89K | 62.5 | 0.3 | |
|
| ||||
| Wildtype | S1-RBD-42 | 42.0,59.0 | 7.5 | |
| Mutant | S1-RBD-42E88K | 64.0 | 0.1 | |
|
| ||||
| LaG24Q68K | Wildtype | S1–14 | 59.5 | 2.4 |
| Mutant | S1–14Q66K | 61.0 | 1.7 | |
|
| ||||
| Wildtype | S1-RBD-31 | 68.5 | 0.2 | |
| Mutant | S1-RBD-31Q67K | 71.5 | 0.4 | |
TM and KD values for wildtype S1–14, S1–28, S1–36 and S1-RBD-14 taken from Mast et al.2
Mutant nanobodies are the wildtype nanobodies incorporating the equivalent original mutation in column 1. (SPR and DSF data detailed in Figures S8 – S9).