Cleavage of fusion proteins using granzyme b protease

C - Chemistry – Metallurgy – 07 – K

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C07K 19/00 (2006.01) C12N 15/62 (2006.01) C12P 21/00 (2006.01) C12P 21/02 (2006.01) C07K 1/12 (2006.01) C12N 9/64 (2006.01)

Patent

CA 2523101

A method for the preparation of a polypeptide of interest in authentic form by enzymatic cleavage of fusion proteins using Granzyme B protease (EC 3.4.21.79). There is also provided fusion proteins comprising a polypeptide of interest and a fusion partner, wherein the junction region between the polypeptide of interest and the fusion partner comprises a Granzyme B protease cleavage site adjacent to the polypeptide of interest, and a human Granzyme B protease variant wherein the Cystein residue no. 228 (chymotrypsinogen numbering) is mutated to Phenylalanine.

L'invention concerne un procédé de préparation d'un polypeptide d'intérêt dans une forme authentique par clivage enzymatique de protéines de fusion au moyen de la protéase granzyme B (EC 3.4.21.79). L'invention concerne également des protéines de fusion comprenant un polypeptide d'intérêt et un partenaire de fusion, la zone de jonction entre le polypeptide d'intérêt et le partenaire de fusion comprenant un site de clivage de la protéase granzyme B jouxtant ledit polypeptide d'intérêt, et une variante de protéase granzyme B, le résidu de cystéine no. 228 (numérotation chymotrypsinogène) mutant vers la phénylalanine.

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