Thromboplastin reagents and methods for preparing and using...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/56 (2006.01) C12N 9/48 (2006.01) C12N 9/64 (2006.01)

Patent

CA 2342606

Disclosed are mammalian thromboplastin reagents and methods for preparing such reagents. The thromboplastin reagents are suitable for use in prothrombin-time (PT) assays, and offer improved sensitivities with acceptable PT-normal times. Contaminating proteins -- particularly plasma clotting factors -- are separated from a thromboplastin solution by a membrane permeation protocol, in which the thromboplastin solution is exposed to a semipermeable membrane and clotting factors are allowed to pass from the thromboplastin solution through the membrane, while Tissue Factor is retained in the thromboplastin solution. In a preferred method, one or more contaminating clotting factors are separated by diafiltration from a NaCl/Na3Citrate thromboplastin extract of mammalian tissue, and the purified thromboplastin extract is combined with Ca++ ions to form a thromboplastin reagent.

La présente invention concerne des réactifs de thromboplastine mammifère et des procédés de leur préparation. Les réactifs de thromboplastine conviennent pour les tests de temps de prothrombine (TP), et présentent des sensibilités améliorées avec des temps normaux de TP. Les protéines contaminantes, en particulier les facteurs de coagulation plasmatique, sont séparées d'une solution de thromboplastine par un protocole de pénétration membranaire, dans lequel on expose la solution de thromboplastine à une membrane semi-perméable et on permet aux facteurs de coagulation de quitter la solution de thromboplastine par la membrane, le facteur tissulaire étant cependant retenu dans la solution de thromboplastine. Dans un procédé préféré, on sépare par diafiltration un ou plusieurs facteurs de coagulation contaminants d'un prélèvement de thromboplastine à citrate NaC1/Na¿3? provenant d'un tissu mammifère, ce prélèvement de thromboplastine purifié étant associé à des ions Ca?++¿ pour former un réactif de thromboplastine.

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