Process for obtaining recombinant prothrombin activating...

C - Chemistry – Metallurgy – 07 – K

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C07K 14/435 (2006.01)

Patent

CA 2577915

This invention refers to the process for obtaining the recombinant prothrombin activating protease (rLopap) in monomeric form, the recombinant prothrombin activating protease (rLopap), as well as its amino acid sequence. In addition to that, this invention also refers to the use of this protease for depleting the blood fibrinogen, and serve as diagnosis kit for dysprothrombinemias. This invention describes the obtainence in recombinant form and the characterization of a prothrombin activator protease of 21 kDa, named rLopap (Lonomia obliqua prothrombin activator protease), with serineproteases characteristics however it shows sequence of conserved amino acids as in a lipocalin family. The protein presents pro-coagulating activity, depleting blood fibrinogen and prolonging the coagulation time of human blood/ plasma. The obtainence of rLopap in its recombinant form and showing adequate activity for allowing clinical Pharmacology essays is presented in this invention.

Cette invention concerne le procédé servant à obtenir une protéase activant la prothrombine recombinée (rLopap) sous une forme monomère, la protéase activant la prothrombine recombinée (rLopap), ainsi que sa séquence d'acides aminés. En plus de cela, cette invention concerne également l'utilisation de cette protéase pour appauvrir le sang en fibrinogène et l'utilisation de celle-ci comme kit de diagnostic pour des dysprothrombinémies. Cette invention décrit l'obtention sous une forme recombinée et la caractérisation d'une protéase activatrice de prothrombine de 21 kDa, appelée rLopap (protéase activatrice de prothrombine du venin de Lonomia obliqua), ayant des caractéristiques de sérineprotéases mais présentant cependant une séquence d'acides aminés conservés comme dans une famille de lipocalines. Cette protéine présente une activité de procoagulation, d'appauvrissement du sang en fibrinogène et de prolongation de la durée de coagulation du sang/plasma humain. L'obtention de la rLopap sous sa forme recombinée et présentant une activité adéquate pour permettre des essais de pharmacologie cliniques est présentée dans cette invention.

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