The goal of the project was to develop a new analytical technique based on activity based enrichment of biological samples containing proteases. The strategy was to bind targeted proteases to inhibitors which are immobilized to a stationary phase, leading to concentration of the targeted proteases on the stationary phase and the possibility to wash away compounds that have no interaction with the affinity ligand.
Controlled Pore Glass beads were used as the stationary phase because of their porosity and high mechanical strength which makes them suitable for use in an HPLC set-up. Purified MMP-12 was used to develop and test the methodology.