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    • 5. 发明申请
    • PLASMA FRACTION PURIFICATION
    • 等离子体分离纯化
    • WO1994001466A1
    • 1994-01-20
    • PCT/US1993006609
    • 1993-07-13
    • ALPHA THERAPEUTIC CORPORATION
    • ALPHA THERAPEUTIC CORPORATIONHERRING, Steven, W.UEMURA, HahiroNODA, MunehiroSHITANISHI, Kenneth, T.
    • C07K15/06
    • C12N9/6429C07K14/745C12N9/6432C12Y304/21005C12Y304/21006
    • The present invention describes a process for activating Factor II to Factor IIa by incubating Factor II in the pressence of Factor V, Factor Xa, phospholipids, and calcium ions. Each of the factors is prepared from a single impure protein fraction which includes Factors II, V and X. The Factor II, V and X purification procedure comprises the steps of DEAE ligand chromatography and precipitation by the addition of barium chloride. Factor V is recovered from the barium chloride supernatant, and Factors II and X are contained in the barium chloride precipitate. The barium chloride precipitate is dissolved in an aqueous solution and is applied to a chromatographic resin coupled with a ligand which binds Factor X and Factor II weakly or not at all. Factor II is recovered from the fraction, which remains unbound or weakly bound to the Factor X binding ligand. Factor Xa is prepared by recovering Factor X from the ligand during the Factor II preparation procedure and activating the Factor X to Factor Xa by specific proteolytic cleavage.
    • 本发明描述了通过在因子V,因子Xa,磷脂和钙离子的压迫下孵育因子II来激活因子II至因子IIa的方法。 每个因素由包含因子II,V和X的单个不纯蛋白质级分制备。因子II,V和X纯化程序包括DEAE配体层析和通过加入氯化钡沉淀的步骤。 因子V从氯化钡上清液中回收,因子II和X包含在氯化钡沉淀中。 将氯化钡沉淀物溶解在水溶液中,并施加到与限制因子X和因子II的配体偶联的色谱树脂上或完全不结合。 因子II从部分回收,其保持未结合或弱结合于因子X结合配体。 因子Xa通过在因子II制备过程中从配体中回收因子X并通过特异性蛋白水解切割活化因子X至因子Xa来制备。