Table 1.
MM kinetics of catalytic Abs
| Entry | Antibody | FRET probe | Km (µM) | kcat (s−1) | kcat/Km (µM−1/s−1) |
| 1 | 18B7 | FRET | 35.8 × 10−3 | 26.4 × 10−3 | 7.3 × 10−1 |
| 2 | 2H1 | FRET | 54.7 × 10−3 | 42.6 × 10-3 | 7.7 × 10−1 |
| 3 | 3E5-IgG1 | FRET | 55.7 × 10−3 | 27.5 × 10−3 | 7.4 × 10−1 |
| 4 | 3E5-IgG3 | FRET | ND | ND | ND |
| 5 | 18B7 | FRET-OAc | 48.3 × 10−3 | 25.7 × 10−3 | 5.3 × 10−1 |
| 6 | 2H1 | FRET-OAc | 36.3 × 10−3 | 33.5 × 10−3 | 9.2 × 10−1 |
| 7 | 3E5-IgG1 | FRET-OAc | 35.9 × 10−3 | 26.7 × 10−3 | 4.9 × 10−1 |
| 8 | 3E5-IgG3 | FRET-OAc | 35.0 × 10−3 | 29.0 × 10−3 | 8.2 × 10−1 |
The initial velocity (V0) of each reaction was determined and plotted as a function of substrate concentration, the data were fit to the MM equation for a single-step bimolecular reaction using nonlinear regression. Km and kcat were calculated using Prism 8. ND, not determined. Each experiment was repeated in triplicate.