C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
C07K 1/113 (2006.01) C07K 14/285 (2006.01) C07K 14/555 (2006.01) C07K 14/575 (2006.01) C07K 16/00 (2006.01) C12N 1/20 (2006.01) C12N 9/48 (2006.01) C12N 9/72 (2006.01) C12N 15/10 (2006.01) C12P 21/00 (2006.01) C12P 21/02 (2006.01) C12P 21/08 (2006.01)
Patent
CA 2313248
A process for the production of a naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges by a) culturing prokaryotic cells in which the said prokaryotic cells contain an expression vector which codes for the said polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) secreting the polypeptide into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium, which is characterized in that a nucleic acid coding for a molecular chaperone is additionally expressed in the said prokaryotic cell and the chaperone is secreted into the periplasm, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.
Divulgation d'un procédé de production d'un polypeptide eucaryotique naturellement replié, contenant plusieurs cystéines liées par des ponts disulfures comprenant les étapes suivantes : a) la culture de cellules procaryotiques contenant un vecteur d'expression codant ledit polypeptide qui contient une séquence signal procaryotique au N-terminal, b) la sécrétion du polypeptide dans le périplasme ou le milieu, c) le clivage de la séquence signal et l'isolement du polypeptide contenu dans le périplasme ou le milieu, qui est caractérisé par le fait qu'un acide nucléique codant un chaperon moléculaire est en plus exprimé dans ladite cellule procaryotique et que le chaperon moléculaire est sécrété dans le périplasme. Ce procédé convient à la production recombinante à haut rendement de polypeptides dans des procaryotes.
Ambrosius Dorothee
Rudolph Rainer
Schaffner Jorg
Schwarz Elisabeth
F. Hoffmann-La Roche Ag
Gowling Lafleur Henderson Llp
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