C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 9/74 (2006.01) C12N 5/10 (2006.01) C12N 15/09 (2006.01)
Patent
CA 2624072
A process for producing a recombinant human thrombin. There is provided a process for producing a recombinant human thrombin, comprising the steps of (1) introducing an expression vector having a gene fragment coding for human prothrombin gene inserted therein in CHO cells to thereby obtain recombinant cells capable of producing human prothrombin; (2) isolating human prothrombin from a culture of the recombinant CHO cells according to anion exchange technique; (3) causing ekaline to act on the isolated human prothrombin to thereby convert the human prothrombin to human thrombin; and (4) isolating human thrombin from the ekaline treatment liquid in accordance with an affinity force based method in which use is made of benzamidine and cation exchange technique. Further, there are provided, obtained by the above process, human thrombin and human prothrombin producing CHO cell.
La présente invention concerne un procédé de production d'une thrombine humaine recombinante qui comprend les étapes suivantes : (1) introduire dans des cellules CHO un vecteur d'expression dans lequel a été inséré un fragment de gène codant pour la prothrombine humaine, afin d'obtenir des cellules recombinantes capables de produire de la prothrombine humaine ; (2) isoler la prothrombine humaine à partir de la culture de cellules CHO recombinantes par une technique d'échange d'anions ; (3) faire agir de l'Ekaline sur la prothrombine humaine isolée afin de convertir la prothrombine humaine en thrombine humaine ; et (4) isoler la thrombine humaine du liquide de traitement à l'Ekaline par un procédé basé sur la force d'affinité qui utilise de la benzamidine et une technique d'échange de cations. L'invention concerne également la thrombine humaine et les cellules CHO produisant de la prothrombine humaine obtenues par le procédé ci-dessus.
Imamura Takayuki
Nakahara Yo
Nakatake Hiroshi
Nouchi Toshinobu
Onchi Tatsufumi
Juridical Foundation The Chemo-Sero-Therapeutic Research Institu
Kirby Eades Gale Baker
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