C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/09 (2006.01) C12N 9/64 (2006.01) C12P 21/02 (2006.01)
Patent
CA 2452768
The present invention is directed to a genetic recombinant human thrombin. Human thrombin is efficiently prepared by a genetic engineering technique comprising the steps: (1) culturing a transfectant animal cell transfected with an expression vector in which a gene encoding human prethrombin is incorporated to the downstream of a promoter so as to produce and accumulate prethrombin in culture supernatant and recovering the produced human prethrombin; (2) treating a solution containing human prethrombin recovered in step (1) with ecarin so as to convert human prethrombin into human thrombin; and (3) purifying the solution obtained after the above activation process to obtain purified human thrombin. The present invention allows for provision of human thrombin on a large scale in a safe and economical manner due to exclusion of blood-derived components.
L'invention concerne de la thrombine génétiquement modifiée, que l'on peut élaborer avec efficacité par modification génique, selon les étapes suivantes: 1) culture de transformant, issu de la transformation d'une cellule animale par un vecteur d'expression porteur de gène codant la préthrombine intégrée en aval d'un promoteur, et collecte de la préthrombine résultante accumulée à l'intérieur du milieu de culture; 2) activation de la solution de préthrombine humaine ainsi recueillie, par traitement à l'écarine, aux fins de conversion en thrombine humaine; et 3) purification de la solution de thrombine humaine résultante, à partir du mélange réactif liquide inactivé comme indiqué précédemment. On peut donc élaborer de la thrombine humaine en quantités importantes, avec une grande sécurité et à faible coût, tout en réduisant au minimum les effets des composantes d'origine sanguine.
Imamura Takayuki
Nakatake Hiroshi
Nozaki Chikateru
Soejima Kenji
Yonemura Hiroshi
Juridical Foundation The Chemo-Sero-Therapeutic Research Institu
Kirby Eades Gale Baker
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