Characterization of TIMP2-MMP binding of the TIMP2 constructs
| Ligand | TIMP2 | TIMP2-hIgG4 | Ala-TIMP2 | |||
|---|---|---|---|---|---|---|
| Binding method | BLI (nm) | SPR (RU) | BLI (nm) | SPR (RU) | BLI (nm) | SPR (RU) |
| Ms-pro-MMP2 | 0.14 | N.D. | 0.016 | N.D. | 0.089 | N.D. |
| Ms-pro-MMP3 | 0.0058 | N.D. | 0.013 | N.D. | 0.013 | N.D. |
| Ms-pro-MMP9 | 0.010 | N.D. | 0.011 | N.D. | 0.0044 | N.D. |
| Hu-pro-MMP2.1 | 0.068 | 5603.6 | 0.017 | 563.87 | 0.070 | 4468.99 |
| Hu-pro-MMP3 | 0.0051 | 337.9 | 0.016 | 249.31 | 0.014 | −85.29 |
| Hu-pro-MMP9 | 0.013 | 219.2 | 0.013 | 164.32 | 0.0076 | 18.63 |
| Hu-pro-MMP2.2 | 0.38 | 1140.07 | 0.079 | 266.57 | 0.37 | 975.75 |
| Hu-CD-MMP3 | 0.015 | 109.39 | 0.035 | 137.61 | 0.025 | 16.64 |
| Hu-CD-MMP9 | 0.019 | 5.03 | 0.025 | 63.44 | 0.021 | 3.10 |
Binding level was analyzed by bio-layer interferometry (BLI) and surface plasmon resonance (SPR) to determine interaction relationships between the TIMP2 constructs and MMP proteins. Binding interactions were assessed for mouse pro- (zymogen) forms and two human forms [pro-, catalytic domain (CD)] of MMP2, MMP3, and MMP9. BLI was performed on Octet Red96e. SPR was performed on Biacore T200. N.D. = No data collected for that combination. N.B. = No binding observed for that combination. Bindings scale cutoffs determined from largest observed binding signal.