Binding kinetics of antibodies in surface plasmon resonance analysis
| Antibody . | Target . | Format . | ka (M−1s−1) . | kd (s−1) . | KD (M) . |
|---|---|---|---|---|---|
| R4-13i | mRobo4 | scFv | 1.25 ± 0.36 × 105 | 5.82 ± 0.95 × 10−4 | 5.03 ± 1.95 × 10−9 |
| (internalizing) | dscFv | 1.15 ± 0.34 × 106 | 5.98 ± 0.61 × 10−4 | 5.64 ± 2.21 × 10−10 | |
| IgG | 1.14 ± 0.55 × 106 | 4.19 ± 1.70 × 10−4 | 2.22 ± 0.51 × 10−10 | ||
| scFv-PSIF | 7.22 ± 4.31 × 104 | 4.28 ± 1.60 × 10−3 | 6.47 ± 1.61 × 10−8 | ||
| IgG-NCS | 1.02 ± 0.15 × 106 | 4.66 ± 0.86 × 10−4 | 4.59 ± 0.74 × 10−10 | ||
| R4-16 | mRobo4 | scFv | 1.30 ± 0.33 × 105 | 5.82 ± 1.50 × 10−4 | 4.77 ± 1.96 × 10−9 |
| (low-internalizing) | dscFv | 1.12 ± 0.03 × 106 | 5.91 ± 1.50 × 10−4 | 5.31 ± 1.96 × 10−10 | |
| IgG | 1.06 ± 0.24 × 106 | 3.60 ± 0.85 × 10−4 | 2.76 ± 0.16 × 10−10 | ||
| scFv-PSIF | 8.90 ± 1.42 × 104 | 6.10 ± 2.45 × 10−3 | 7.24 ± 3.74 × 10−8 | ||
| IgG-NCS | 1.07 ± 0.12 × 106 | 3.93 ± 0.54 × 10−4 | 3.72 ± 0.89 × 10−10 | ||
| V2-05i | mVEGFR2 | scFv | 9.66 ± 3.57 × 104 | 4.40 ± 0.95 × 10−4 | 5.13 ± 2.61 × 10−9 |
| (internalizing) | dscFv | 8.75 ± 2.03 × 105 | 5.59 ± 2.57 × 10−4 | 6.16 ± 1.47 × 10−10 | |
| IgG | 1.14 ± 0.09 × 106 | 3.21 ± 0.35 × 10−4 | 2.84 ± 0.52 × 10−10 | ||
| scFv-PSIF | 9.57 ± 0.84 × 104 | 6.51 ± 1.87 × 10−3 | 6.94 ± 2.63 × 10−8 | ||
| IgG-NCS | 0.96 ± 0.06 × 106 | 4.37 ± 0.90 × 10−4 | 4.52 ± 0.79 × 10−10 | ||
| V2-02 | mVEGFR2 | scFv | 7.94 ± 1.24 × 104 | 4.28 ± 3.23 × 10−4 | 5.07 ± 3.05 × 10−9 |
| (low-internalizing) | dscFv | 8.94 ± 2.55 × 105 | 5.57 ± 1.25 × 10−4 | 6.60 ± 2.39 × 10−10 | |
| IgG | 1.13 ± 0.22 × 106 | 3.25 ± 1.10 × 10−4 | 2.90 ± 0.98 × 10−10 | ||
| scFv-PSIF | 9.84 ± 1.52 × 104 | 5.75 ± 2.05 × 10−3 | 5.81 ± 1.93 × 10−8 | ||
| IgG-NCS | 1.08 ± 0.08 × 106 | 5.25 ± 1.58 × 10−4 | 4.85 ± 1.30 × 10−10 |
| Antibody . | Target . | Format . | ka (M−1s−1) . | kd (s−1) . | KD (M) . |
|---|---|---|---|---|---|
| R4-13i | mRobo4 | scFv | 1.25 ± 0.36 × 105 | 5.82 ± 0.95 × 10−4 | 5.03 ± 1.95 × 10−9 |
| (internalizing) | dscFv | 1.15 ± 0.34 × 106 | 5.98 ± 0.61 × 10−4 | 5.64 ± 2.21 × 10−10 | |
| IgG | 1.14 ± 0.55 × 106 | 4.19 ± 1.70 × 10−4 | 2.22 ± 0.51 × 10−10 | ||
| scFv-PSIF | 7.22 ± 4.31 × 104 | 4.28 ± 1.60 × 10−3 | 6.47 ± 1.61 × 10−8 | ||
| IgG-NCS | 1.02 ± 0.15 × 106 | 4.66 ± 0.86 × 10−4 | 4.59 ± 0.74 × 10−10 | ||
| R4-16 | mRobo4 | scFv | 1.30 ± 0.33 × 105 | 5.82 ± 1.50 × 10−4 | 4.77 ± 1.96 × 10−9 |
| (low-internalizing) | dscFv | 1.12 ± 0.03 × 106 | 5.91 ± 1.50 × 10−4 | 5.31 ± 1.96 × 10−10 | |
| IgG | 1.06 ± 0.24 × 106 | 3.60 ± 0.85 × 10−4 | 2.76 ± 0.16 × 10−10 | ||
| scFv-PSIF | 8.90 ± 1.42 × 104 | 6.10 ± 2.45 × 10−3 | 7.24 ± 3.74 × 10−8 | ||
| IgG-NCS | 1.07 ± 0.12 × 106 | 3.93 ± 0.54 × 10−4 | 3.72 ± 0.89 × 10−10 | ||
| V2-05i | mVEGFR2 | scFv | 9.66 ± 3.57 × 104 | 4.40 ± 0.95 × 10−4 | 5.13 ± 2.61 × 10−9 |
| (internalizing) | dscFv | 8.75 ± 2.03 × 105 | 5.59 ± 2.57 × 10−4 | 6.16 ± 1.47 × 10−10 | |
| IgG | 1.14 ± 0.09 × 106 | 3.21 ± 0.35 × 10−4 | 2.84 ± 0.52 × 10−10 | ||
| scFv-PSIF | 9.57 ± 0.84 × 104 | 6.51 ± 1.87 × 10−3 | 6.94 ± 2.63 × 10−8 | ||
| IgG-NCS | 0.96 ± 0.06 × 106 | 4.37 ± 0.90 × 10−4 | 4.52 ± 0.79 × 10−10 | ||
| V2-02 | mVEGFR2 | scFv | 7.94 ± 1.24 × 104 | 4.28 ± 3.23 × 10−4 | 5.07 ± 3.05 × 10−9 |
| (low-internalizing) | dscFv | 8.94 ± 2.55 × 105 | 5.57 ± 1.25 × 10−4 | 6.60 ± 2.39 × 10−10 | |
| IgG | 1.13 ± 0.22 × 106 | 3.25 ± 1.10 × 10−4 | 2.90 ± 0.98 × 10−10 | ||
| scFv-PSIF | 9.84 ± 1.52 × 104 | 5.75 ± 2.05 × 10−3 | 5.81 ± 1.93 × 10−8 | ||
| IgG-NCS | 1.08 ± 0.08 × 106 | 5.25 ± 1.58 × 10−4 | 4.85 ± 1.30 × 10−10 |
Binding kinetics were analyzed against mRobo4 (R4-13i and R4-16) or mVEGFR2 (V2-05i and V2-02). Values are shown as means ± SD from three different preparations.
ka, association rate constant (M−1s−1); kd, dissociation rate constant (s−1); KD, equilibrium dissociation constant (kd/ka) (M).