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ENDOTOXIN-INDUCED THROMBOSIS FACTOR WHICH INDUCES PROCOAGULANT ACTIVITY IN EPITHELIAL CELLS

机译:内毒素诱导的上皮细胞血栓形成因子

摘要

This invention provides a purified endotoxin-induced thrombosis factor, preferably an endotoxin-induced thrombosis factor characterized by an apparent molecular weight between about 50,000 and 65,000 daltons, more specifically about 55,000 daltons, on reduced and nonreduced SDS-polyacrylamide gels, by maximal recovery on elution from such gels at 52,000 to 58,000 daltons, by the ability to migrate as a single band on such gels, by the ability to precipitate in ammonium sulfate at saturations from 40% to 70%, by the ability to precipitate in polyethylene glycol at concentrations above 15%, by high hydrophobicity, by the ability to bind weakly to a hydroxylapatite column and to a lentil lectin column, by the ability to bind tightly to a hydrophobic interaction resin and smear off with ethylene glycol, and by the ability to bind tightly to a reverse-phase column and elute more effectively with isopropranol than with acetonitrile, by the ability to bind to an anion exchange resin over a pH range from 5 to 10, by the inability to bind to a cation exchange resin, by resistance to acid denaturation up to 30 minutes, resistance to polymyxin, sensitivity to heating at 95 DEG C. for 30 minutes, and sensitivity to trypsin exposure for 24 hours. Another characteristic of a purified endotoxin-induced thrombosis factor that it maximally induces tissue factor after six to eight hours, and continues to induce tissue factor for up to sixty hours. This invention also provides purified nucleic acid molecules, antibodies, an inhibitor, an antagonist, pharmaceutical compositions, methods of treatment, and methods of preparation all directed to endotoxin-induced thrombosis factor.
机译:本发明提供了纯化的内毒素诱导的血栓形成因子,优选内毒素诱导的血栓形成因子,其特征在于在还原的和未还原的SDS-聚丙烯酰胺凝胶上通过最大回收率在约50,000-65,000道尔顿,更具体地约55,000道尔顿之间的表观分子量。通过在此类凝胶上以单条带迁移的能力,通过在硫酸铵中以40%至70%的饱和度沉淀的能力,通过在聚乙二醇中以一定浓度沉淀的能力,从这类凝胶中洗脱52,000至58,000道尔顿高于15%,是由于高疏水性,与羟基磷灰石色谱柱和小扁豆凝集素柱弱结合的能力,与疏水相互作用树脂紧密结合并被乙二醇涂抹的能力以及与乙二醇紧密结合的能力通过在pH值下与阴离子交换树脂结合的能力,将其连接到反相柱上,并用异丙醇比用乙腈更有效地洗脱不能结合阳离子交换树脂,耐酸变性长达30分钟,抗多粘菌素,对95℃加热30分钟的敏感性以及对胰蛋白酶暴露24的敏感性,范围为5至10小时。纯化的内毒素诱导的血栓形成因子的另一个特征是,它在六到八小时后最大程度地诱导组织因子,并在长达六十小时的时间内持续诱导组织因子。本发明还提供了全部针对内毒素诱导的血栓形成因子的纯化的核酸分子,抗体,抑制剂,拮抗剂,药物组合物,治疗方法和制备方法。

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