Table 1.
Affinity determination against human and mouse EGFR.
| Human EGFR | Mouse EGFR | |||||||
|---|---|---|---|---|---|---|---|---|
| KD (nM) | kon (106 M−1.s−1) | koff (10−2 s−1) | R² | KD (nM) | kon (106 M−1.s−1) | koff (10−2 s−1) | R² | |
| D8 | 5.56 ± 0.05 | 1.55 ± 0.01 | 0.86 ± 0.00 | 0.96 | 59.69 ± 3.01 | 1.74 ± 0.08 | 10.40 ± 0.16 | 0.94 |
| D8-HA | 5.29 ± 0.05 | 1.70 ± 0.02 | 0.90 ± 0.00 | 0.96 | 54.17 ± 2.61 | 1.77 ± 0.08 | 9.59 ± 0.14 | 0.94 |
| D8-vc-MMAE | 5.85 ± 0.08 | 3.88 ± 0.05 | 2.27 ± 0.01 | 0.99 | N.D. | N.D. | N.D. | N.D. |
| B10a | 27.6 | 1.90 | 5.24 | 0.99 | 83.0 | 1.41 | 11.7 | 0.98 |
| B10-HA | 48.80 ± 1.39 | 2.09 ± 0.06 | 10.18 ± 0.09 | 0.98 | 143.60 ± 5.92 | 2.68 ± 0.10 | 38.44 ± 0.53 | 0.99 |
| B10-vc-MMAE | 114.20 ± 6.40 | 2.03 ± 0.11 | 23.14 ± 0.39 | 0.98 | N.D. | N.D. | N.D. | N.D. |
Binding characteristics of the antiEGFR Nanofitins and Nanofitin-drug conjugates determined by biolayer interferometry on human and mouse EGFR. D8 and B10: Nanofitins without C-terminal tag. D8-HA and B10-HA: Nanofitins with a C-terminal HA-tag. D8-vc-MMAE and B10-vc-MMAE: Nanofitin-drug conjugates with a C-terminal payload.
aAccording to Goux, et al (32). N.D.: Not determined.