Characterization of TIMP2 protein constructs
| Construct | Identity (intact mass) | Concentration (mg/ml; A280) | Purity (%; R/NR, SDS-PAGE) | Aggregation (SEC-MALS) | Oligomerization (SEC-MALS) | Endotoxin (EU/ml; LAL) | |
|---|---|---|---|---|---|---|---|
| TIMP2 | Confirmed | 1.57 | 82/100 | None detected | Monomer | <0.1 | |
| TIMP2-hIgG4 | Confirmed | 1.89 | 100/85 | <5% | Dimer | <0.1 | |
| Ala-TIMP2 | Confirmed | 2.47 | 75–81/100 | None detected | Monomer | <0.1 | |
All recombinant TIMP2 protein constructs were characterized for identity (intact mass), concentration [A280, adjusted with calculated extinction coefficient: TIMP2, Ala-TIMP2 Abs 0.1% (=1 g/l) = 1.525; TIMP2-hIgG4 Abs 0.1% (= 1 g/l) = 1.408)], purity (SDS-PAGE), oligomerization and aggregation (SEC-MALS), and endotoxin (LAL assay) characterization. “/” in Purity column used to separate Reduced and Nonreduced percentage values, not division. R, reducing; NR, non-reducing; EU, endotoxin unit; LAL, limulus amebocyte lysate; SEC, size exclusion chromatography; MALS, multi angle light scattering.