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Purification of plasma fraction

机译:血浆馏分的纯化

摘要

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.
机译:本发明描述了通过在因子V,因子Xa,磷脂和钙离子的压力下温育因子II将因子II活化为因子IIa的方法。每种因子均由单个不纯的蛋白级分制备,其中包括因子II,V和X。因子II,V和X纯化程序包括DEAE配体色谱法步骤和通过添加氯化钡进行沉淀的步骤。从氯化钡上清液中回收因子V,氯化钡沉淀中包含因子II和X。将氯化钡沉淀物溶解在水溶液中,并施加到与配体偶联的色谱树脂上,该配体与因子X和因子II的结合弱或根本不结合。从级分中回收因子II,该级分仍未与因子X结合配体结合或弱结合。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裂解将因子X激活为因子Xa来制备因子Xa。

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