首页> 外国专利> FRACTION OF LYSINE-RICH HISTONE THYMUS FACTOR (TF), METHOD FOR PREPARING THE FRACTION AND METHOD FOR DETECTING HUMAN ACQUIRED IMMUNODEFIENCY SYNDROME

FRACTION OF LYSINE-RICH HISTONE THYMUS FACTOR (TF), METHOD FOR PREPARING THE FRACTION AND METHOD FOR DETECTING HUMAN ACQUIRED IMMUNODEFIENCY SYNDROME

机译:富含赖氨酸的组蛋白胸腺因子(TF)的成分,制备方法和检测人源性免疫缺陷综合症的方法

摘要

1. A fraction of lysine-rich histone thymus factor (TF) of mammals, capable of precipitating when contacted with serum of a human, uninfected with HIV, forming one continuous precipitation spur on the gel after two-dimensional electrophoresis in relation to the location of -, 1- and 2- macroglobulins, and not capable of precipitating when contacted with serum of a human, infected with HIV, forming one continuous precipitation arch on the gel after two-dimensional electrophoresis in relation to the location of -, 1- and 2- macroglobulins. 2. A TF fraction of claim 1, wherein, when methylated or unmethylated, has a pH of between 7.64 and 8.6, a molecular weight of methylated TF is about 28 kilodalton as determined by 11% non-reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a molecular weight of unmethylated TF is about 35 kilodalton as determined by 11% non-reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and has an isoelectric point of about 5.6. 3. A method of obtaining TF fraction of claim 1, comprising extracting a lysine-rich histone fraction from mammalian thymus cell nuclei by means of pepsin, contacting the lysine-rich histone fraction with a cation exchange chromatographic material, eluting the TF protein with an effective aqueous salt solution and collecting the obtained fraction. 4. A method of claim 3, wherein the cation exchange chromatography is performed on a carboxymethyl column and the TF protein is eluted by applying a linear gradient of sodium chloride at pH 6.8. 5. A method for detecting acquired immunodeficiency syndrome in a human, comprising collecting a biological sample from the subject, suspected for HIV, providing contacting of said sample with the fraction of claim 1, correlating precipitation patterns TF and aTF in a tested sample with the precipitation pattern TF and aTF obtained from a HIV infected human, by detecting localization products of precipitation respective to the zone of the location of -, 1- and 2- macroglobulins. wherein the absence of the spur precipitaion or the presence of the broken precipitation spur within or beyond the location of -, 1- and 2- macroglobulins proves the presence of HIV infection in a human.
机译:1.一部分富含赖氨酸的哺乳动物组蛋白胸腺因子(TF),当与未感染HIV的人的血清接触时能够沉淀,在二维电泳后与位置相关的凝胶上形成一个连续的沉淀刺-,1-和2-巨球蛋白,当与感染HIV的人血清接触时不能沉淀,在二维电泳后,相对于-,1-的位置在凝胶上形成一个连续的沉淀拱和2个巨球蛋白。 2.权利要求1的TF级分,其中当甲基化或未甲基化时,其pH在7.64至8.6之间,通过11%的非还原十二烷基硫酸钠-聚丙烯酰胺凝胶电泳确定,甲基化TF的分子量为约28千道尔顿。通过11%的非还原十二烷基硫酸钠-聚丙烯酰胺凝胶电泳确定,未甲基化的TF的分子量为约35千道尔顿,等电点为约5.6。 3.权利要求1的获得TF级分的方法,其包括通过胃蛋白酶从哺乳动物胸腺细胞核中提取富含赖氨酸的组蛋白级分,使富含赖氨酸的组蛋白级分与阳离子交换层析材料接触,用TFA洗脱TF蛋白。有效的盐水溶液并收集得到的级分。 4.根据权利要求3所述的方法,其中所述阳离子交换色谱法在羧甲基柱上进行,并且所述TF蛋白通过在pH 6.8下施加氯化钠的线性梯度来洗脱。 5.一种用于检测人中获得性免疫缺陷综合症的方法,其包括从疑似HIV的受试者中收集生物样品,使所述样品与权利要求1的级分接触,使测试样品中的沉淀模式TF和aTF与所述样品的沉淀模式相关。通过检测分别对应于-,1-和2-大球蛋白位置区域的降水的定位产物,从HIV感染者中获得TF和aTF的沉淀模式。其中在-,1-和2-巨球蛋白的位置之内或之外不存在刺突沉淀或破裂的沉淀刺突的存在证明了人类中存在HIV感染。

著录项

  • 公开/公告号EA001100B1

    专利类型

  • 公开/公告日2000-10-30

    原文格式PDF

  • 申请/专利权人 TOMPSON U.S.A. LTD.;

    申请/专利号EA19970000353

  • 发明设计人 ZHABILOV HARRY P.;

    申请日1996-05-01

  • 分类号C07K14/47;G01N33/569;A61K38/17;A61K39/00;A61P31/18;

  • 国家 EA

  • 入库时间 2022-08-22 01:56:37

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