Table 1.
Equilibrium KD (KD in nanomolar, nM) for binding of the set of 10 Nbs (belonging to five functional groups based on their effect on kinase activity) to either FL-LRRK2 or its RCKW domain construct, as assessed by two methods in parallel: MST and BLI
| Nb | Binding epitope (ELISA) | Nb group | FL-LRRK2 KD (nM) | RCKW KD (nM) | |
| MST | BLI | BLI | |||
| Nb1 | COR-B | Group 1 | 91 ± 28 | 55 ± 12 | 39 ± 4 |
| Nb6 | COR-B | Group 1 | 83 ± 24 | 8 ± 1 | 5 ± 1 |
| Nb23 | K-WD40 | Group 1 | NBS* | 20 ± 5 | 17 ± 7 |
| Nb17 | FL-LRRK2‡ | Group 2 | 67 ± 24 | 2.0 ± 0.3 | NB† |
| Nb36 | FL-LRRK2‡ | Group 2 | 78 ± 25 | 16 ± 8 | NB† |
| Nb38 | FL-LRRK2‡ | Group 2 | 48 ± 11 | 37 ± 4 | NB† |
| Nb22 | K-WD40 | Group 3 | 145 ± 55 | 10 ± 1 | 5 ± 1 |
| Nb42 | Roc | Group 4 | 94 ± 30§ | 58 ± 18 | 18 ± 2 |
| Nb39 | FL-LRRK2‡ | Group 5 | 79 ± 19 | 12 ± 3 | NB† |
| Nb40 | K-WD40 | Group 5 | 26 ± 10 | 2.5 ± 0.3 | 2.6 ± 0.3 |
*NBS: no binding signal observed in MST.
†NB: no binding on RCKW.
‡FL-LRRK2: full-length LRRK2.
§Value determined in presence of GTPγS instead of GDP, which was used in all the other measurements.