C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/29 (2006.01) C07K 14/81 (2006.01) C12N 9/50 (2006.01) C12N 9/72 (2006.01)
Patent
CA 2185436
Process for purifying serine proteases from a protein mixture by binding the serine protease to an immobilized polypeptide with the activity of an inhibitor DE-3 from Erythrina caffra, removing unbound components from the protein mixture, detaching the serine protease from the inhibitor and separating the immobilized inhibitor from the soluble serine protease and isolating serine protease which is characterized in that a polypeptide is used as the polypeptide which is the product of a prokaryotic or eukaryotic expression of an exogenous nucleic acid. This inhibitor is distinguished by an improved specific activity and is particularly suitable for the purification of plasminogen activators such as tissue plasminogen activators (t-PA and derivatives).
L'invention concerne un procédé de purification de sérine protéases provenant d'un mélange de protéines, qui consiste à lier la sérine protéase à un polypeptide immobilisé ayant l'activité d'un inhibiteur DE-3 issu de l'Erythrina caffra, à extraire des constituants non liés du mélange de protéines, à séparer la sérine protéase de l'inhibiteur et à séparer l'inhibiteur immobilisé de la sérine protéase soluble, ainsi qu'à isoler la sérine protéase. Ce procédé se caractérise en ce que le polypeptide utilisé est un polypeptide qui est le produit de l'expression procaryote ou eucaryote d'un acide nucléique exogène. Cet inhibiteur se caractérise par une meilleure activité spécifique et s'utilise notamment dans l'épuration d'activateurs du plasminogène tels que des activateurs tissulaires du plasminogène (t-PA et dérivés).
Fischer Stephan
Kohnert Ulrich
Stern Anne
Norton Rose Or S.e.n.c.r.l. S.r.l./llp
Roche Diagnostics Gmbh
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