Production of multimeric fusion proteins using a c4bp scaffold

Details Production of multimeric fusion proteins using a c4bp scaffold Production of multimeric fusion proteins using a c4bp scaffold

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) A61K 38/17 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/63 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2494981

The present invention provides a method for obtaining a recombinant fusion protein comprising a scaffold of a C-terminal core protein of C4bp alpha chain, said recombinant fusion protein being capable of forming multimers in soluble form in a prokaryotic host cell, the method including the steps of (i) providing a prokaryotic host cell carrying a nucleicacid encoding said recombinant protein operably linked to a promoter functional in said prokaryotic cell; (ii) culturing the host cell under conditions whereinsaid recombinant protein is expressed; and (iii) recovering the recombinant protein wherein said protein is recovered in multimeric form without performing a scaffold refolding step.

La présente invention concerne un procédé d'obtention d'une protéine de fusion recombinante comprenant un échafaudage d'une apoenzyme à terminaison C d'une chaîne alpha C4BP, ladite protéine de fusion recombinante étant capable de former des multimères sous forme soluble dans une cellule hôte procaryote, le procédé consistant à (i) apporter une cellule hôte procaryote portant un acide nucléique codant ladite protéine recombinante liée de manière opérationnelle à une promoteur fonctionnel dans ladite cellule procaryote ; (ii) cultiver la cellule hôte dans les conditions dans lesquelles ladite protéine recombinante est exprimée ; et (iii) récupérer la protéine recombinante sous forme multimère sans réaliser de repliage de protéine d'échafaudage.

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