AN ARTIFICIAL 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 vitrotestingsystems containing a group of structural elements comprising the signalsequence of CD5, the FLAG-epitope for M1 monoclonal antibody detection, theinterglobular domain of human aggrecan, the hinge region of human IgG1, theCH2 region of human IgG1 and the CH3 region of human IgG1; a DNAencoding the recombinant substrate; vectors and host cells containing said DNAand use of the recombinant substrate to study the activity of aggrecanase todetect new enzymatic cleavage sites, to purify aggrecanasechromatographically, to clone the aggrecanase cDNA by functional cloning, toscreen for an aggrecanase inhibitor; a method for monitoring the onset orprogression of osteorthritis and a diagnostic aid containing the recombinantsubstrate.