An artifical recombinant substrate (rAGG-1) and native aggrecan to study the proteolytic activity of 'Aggrecanase' in cell culture systems

摘要

The invention refers to a recombinant substrate for Aggrecanase in vitro testing systems containing a group of structural elements comprising the signal sequence of CD5, the FLAG-epitope for M1 monoclonal antibody detection, the interglobular domain of human aggrecan, the hinge region of human IgG1, the CH2 region of human IgG1 and the CH3 region of human IgG1; a DNA encoding the recombinant substrate; vectors and host cells containing said DNA and use of the recombinant substrate to study the activity of aggrecanase to detect new enzymatic cleavage sites, to purify aggrecanase chromatographically, to clone the aggrecanase cDNA by functional cloning, to screen for an aggrecanase inhibitor; a method for monitoring the onset or progression of osteorthritis and a diagnostic aid containing the recombinant substrate.