公开(公告)号：US5102807A

公开(公告)日：1992-04-07

申请号：US336052

申请日：1989-04-11

申请人： Henry G. Burger ,  David M. de Kretser ,  John K. Findlay ,  Francis J. Morgan ,  Milton T. W. Hearn ,  David Milne-Robertson ,  Robert G. Forage ,  Richard E. H. Weitenhall

发明人： Henry G. Burger ,  David M. de Kretser ,  John K. Findlay ,  Francis J. Morgan ,  Milton T. W. Hearn ,  David Milne-Robertson ,  Robert G. Forage ,  Richard E. H. Weitenhall

IPC分类号： C07K14/435 ,  A61K35/48 ,  A61K35/54 ,  A61K38/00 ,  A61K38/22 ,  A61K39/00 ,  A61K39/395 ,  C07K1/16 ,  C07K1/20 ,  C07K1/26 ,  C07K14/575 ,  C07K16/00 ,  C07K16/26 ,  G01N33/68

CPC分类号： G01N33/689 ,  C07K14/575 ,  C07K16/26 ,  A61K38/00 ,  A61K39/00 ,  Y10S436/817 ,  Y10S530/806 ,  Y10S530/85

摘要： A protein which satisfies all the biological criteria which are characteristic of inhibin has been isolated from a gonadal source. The purification and characterization of inhibin and the use of the purified material to raise antibodies, the use of inhibin and said antisera in a quantative radioimmunoassay, and applications in vitro and in vivo of inhibin and antibody to inhibin, are described.There is provided a purified protein, inhibin, characterised in thata. the apparent molecular weight as determined by SDS-PAGE is 56,000.+-.1,000b. the isoelectric point is in the range 6.9-7.3c. the protein can bind specifically to Concanavalin A-Sepharosed the protein consists of two sub-units, characterized in thati. their apparent molecular weights as determined by SDS-PAGE are 44,000.+-.3,000 and 14,000.+-.2,000 respectively.ii. the isoelectric point of the 44,000 molecular weight sub-unit is in the range 6.0-7.0iii. the N-terminal amino acid sequences of the two sub-units are as described hereine. the protein can suppress follicle stimulating hormone (FSH) but not luteinising hormone (LH), thyroid stimulating hormone or prolactin in an in vitro bioassay systemf. the protein can be labeled with radioactive iodine.There is also provided a method for isolating and purifying inhibin from mammalian ovarian follicular fluid, characterised bya. one or more gel permeation chromatography steps;b. one or more reversed-phase high performance liquid chromatography steps;c. one or more preparative polyacrylamide gel electrophoresis steps;d. Electrophoretic elution of the purified inhibin.

摘要翻译： 已经从性腺源分离了满足抑制素特征的所有生物学标准的蛋白质。 纯化和抑制素的表征和使用该纯化的物质的产生抗体，使用抑制素和所述一个定量放射免疫测定的抗血清，并在体外和抑制素和抗体抑制素的体内应用，进行了描述。 提供了一种纯化的蛋白质抑制素，其特征在于a。 通过SDS-PAGE测定的表观分子量为56,000 +/- 1,000b。 等电点在6.9-7.3 c的范围内。 蛋白质可以特异性结合伴刀豆球蛋白A-Sepharose d，蛋白质由两个亚单位组成，其特征在于i。 它们通过SDS-PAGE测定的表观分子量分别为44,000 +/- 3,000和14,000 +/- 2,000。 ii。 44,000分子量子单元的等电点在6.0-7.0 iii范围内。 两个亚单位的N-末端氨基酸序列如本文所述。 该蛋白质能够抑制促卵泡激素（FSH），但不黄体生成激素（LH），促甲状腺激素或促乳素在体外生物测定系统F。 蛋白质可以用放射性碘标记。 还提供了一种从哺乳动物卵巢滤泡液中分离和纯化抑制素的方法，其特征在于a。 一个或多个凝胶渗透色谱步骤; b。 一个或多个反相高效液相色谱步骤; C。 一个或多个制备型聚丙烯酰胺凝胶电泳步骤; d。 纯化的抑制素的电泳洗脱。

公开(公告)号：US5364837A

公开(公告)日：1994-11-15

申请号：US389328

申请日：1989-08-03

申请人： Henry G. Burger ,  David M. de Kretser ,  John K. Findlay ,  Francis J. Morgan ,  Milton T. W. Hearn ,  David M. Milne-Robertson ,  Robert G. Forage

发明人： Henry G. Burger ,  David M. de Kretser ,  John K. Findlay ,  Francis J. Morgan ,  Milton T. W. Hearn ,  David M. Milne-Robertson ,  Robert G. Forage

IPC分类号： C07K14/435 ,  A61K35/48 ,  A61K35/54 ,  A61K38/00 ,  A61K38/22 ,  A61K39/00 ,  A61K39/395 ,  C07K1/16 ,  C07K1/20 ,  C07K1/26 ,  C07K14/575 ,  C07K16/00 ,  C07K16/26 ,  G01N33/68 ,  A61K37/24

CPC分类号： G01N33/689 ,  C07K14/575 ,  C07K16/26 ,  A61K38/00 ,  A61K39/00 ,  Y10S436/817 ,  Y10S530/806 ,  Y10S530/85

摘要： A protein which satisfies all the biological criteria which are characteristic of inhibin has been isolated from a gonadal source. The purification and characterization of inhibin and the use of the purified material to raise antibodies, the use of inhibin and said antisera in a quantative radioimmunoassay, and application in vitro and in vivo of inhibin and antibody to inhibin, are described. There is provided a purified protein, inhibin characterized in that the apparent molecular weight as determined by SDS-PAGE is 56,000.+-.1,000, the isoelectric point is in the range 6.9-7.3, and the protein can bind specifically to Concanavalin A-Sepharose. Moreover, the protein includes two subunits, characterized in that their apparent molecular weights as determined by SDS-PAGE are 44,000.+-.3,000 and 14,000.+-.2,000, respectively. Furthermore, the isoelectric point of the 44,000 molecular weight sub-unit is in the range of 6.0-7.0. In addition, the N-terminal amino acid sequence of the two subunits are described. With regard to the purified protein, inhibin, the protein can suppress follicle stimulating hormone (FSH) but not luteinizing hormone (EACH), thyroid stimulating hormone or prolactin in an in vitro bioassay system, and the protein can be labeled with radioactive iodine. In addition to the forgoing, there is also provided a method for isolating and purifying inhibin from mammalian ovarian follicular fluid, characterized by one or more gel permeation chromatography steps; reversed-phase high performance liquid chromatography steps; preparative polyacrylamide gel electrophoresis steps; and electrophoretic elution of the purified inhibin.

摘要翻译： 已经从性腺源分离了满足抑制素特征的所有生物学标准的蛋白质。 描述了抑制素的纯化和表征以及纯化材料用于产生抗体的用途，在定量放射免疫测定中使用抑制素和所述抗血清，以及体外和体内抑制素和抗体对抑制素的应用。 提供了纯化的蛋白质，抑制素，其特征在于通过SDS-PAGE测定的表观分子量为56,000 +/- 1,000，等电点在6.9-7.3范围内，蛋白质可以特异性结合伴刀豆球蛋白A-琼脂糖凝胶 。 此外，蛋白质包括两个亚基，其特征在于，其通过SDS-PAGE测定的表观分子量分别为44,000 +/- 3,000和14,000 +/- 2,000。 此外，44,000分子量子单元的等电点在6.0-7.0的范围内。 此外，描述了两个亚基的N-末端氨基酸序列。 关于纯化的蛋白质，抑制素，蛋白质可以在体外生物测定系统中抑制卵泡刺激素（FSH），而不是黄体生成激素（EACH），促甲状腺激素或催乳素，并且蛋白质可以用放射性碘标记。 除了上述之外，还提供了分离和纯化哺乳动物卵泡滤液中抑制素的方法，其特征在于一个或多个凝胶渗透色谱步骤; 反相高效液相色谱步骤; 制备聚丙烯酰胺凝胶电泳步骤; 和纯化的抑制素的电泳洗脱。