An artificial recombinant substrate (ragg 1) and native...

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/62 (2006.01) C07K 14/47 (2006.01) C07K 16/00 (2006.01) C07K 19/00 (2006.01) C12N 9/64 (2006.01) C12N 15/10 (2006.01) C12N 15/12 (2006.01) C12Q 1/37 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2195385

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.

a présente invention porte sur un substrat recombinant pour des systèmes d'essai in vitro d'une Aggrecanase renfermant un groupe d'éléments structuraux comprenant la séquence-signal de CD5, l'épitope-FLAG pour le dépistage des anticorps monoclonaux M1, le domaine interglobulaire de l'aggrecan humaine, la partie charnière d'IgG1 humaine, la région CH2 d'IgG1 humaine et la région CH3 d'IgG1 humaine; un ADN codant pour le substrat recombinant; des vecteurs et des cellules-hôtes renfermant le dit ADN et l'utilisation du substrat recombinant pour étudier l'activité de l'aggrecanase pour déceler de nouveaux sites de coupure enzymatique, pour purifier l'aggrecanase par chromatographie, pour cloner l'ADNc de l'aggrecanase par clonage fonctionnel, pour rechercher un inhibiteur de l'aggrecanase; une méthode pour surveiller l'apparition ou l'évolution de l'ostéo-arthrite et un moyen de diagnostic renfermant le substrat recombinant.

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