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Development of the ultra sensitive sandwich assay of human inhibin and it's clinical applocations

Research Project

Project/Area Number 08557091
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Obstetrics and gynecology
Research InstitutionGunma University

Principal Investigator

IBUKI Yoshito  Gunma Univ.Sch.Med.Dept.OB/GIN., Prof., 医学部, 教授 (40008256)

Co-Investigator(Kenkyū-buntansha) MASHIMO Ikuo  SRL,Inc.Res.Div., Associate Director, 研究検査部, 担当次長研究員
HASEGAWA Yoshihisa  Kitasato Univ.Suh.Vet.Med.Anim.Sci., Prof., 獣医畜産学部, 教授 (40092001)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1996: ¥10,300,000 (Direct Cost: ¥10,300,000)
KeywordsInhibin / Sandwich Assay / Clinical Applications / アッセイ法 / 高感度化
Research Abstract

Inhibin shows molecular heterogeneity and there exist not only biologically active dimers but also immunologically active and biologically inactive monomers in human body fluid. It is also widely recognized that several proteins including follistatin and alpha 2 macroglobulin binding to inhibin and structurally related activin interfere with the assay of inhibin.
In order to establish a highly sensitive and specific sandwich assay of biologically active inhibin, we investigated the character of human inhibin, produced a variety of antibodies against inhibin and studied several methods of labeling the antibodies.
First we established a large scale purification system for inhibin from human follicular fluid. It was then possible to obtain the required amount of inhibin for a standard in the assay kit. Recombinant human inhibin B was expressed, purified and characterized and we confirmed that this recombinant inhibin is useful as an inhibin B assay standard (Hasegawa et al.J Reprod Develop … More 42 : 23-25,1996. Inhibin and Activin pp139-154,1996. Hasegawa et al. Sexual Differentiation and Maturation pp139-154,1996).
Next, we investigated labeling methods using many kinds of polyclonal and monoclonal antibodies. Using an anti alpha subunit polyclonal antibody coated on a bead and anti beta_A subunit monoclonal antibody labeled by ^<125>I,we established an inhibin A radioimmunometric assay (Inhibin A IRMA) (Hasegawa et al.Inhibin, Activin, and Follistatin, pp104-117,1997). Changes in inhibin A concentrations in circulating blood during normal human pregnancy and abnormal pregnancy such as pregnancy induced hypertension and intrauterine growth retardation were clarified using this inhibin A IRMA (Mizunuma et al. Inhibin, Activin, and Follistatin pp151-161,1997). Because this inhibin A IRMA is not sensitive enough to measure the inhibin concentrations in circulating blood in non-pregnant women, further investigation has been done in order to improve its sensitivity. The polyclonal antibody was purified by an inhibin coupled column and an activin coupled column and thus an anti alpha subunit antibody and an anti beta_A subunit antibody were obtained. Inhibin A immunofluorometric assay (Inhibin A IFMA) was established using microtiter plates coated with the anti beta_A subunit antibody and the anti alpha subunit antibody labeled with Europium. This inhibin A IFMA is sensitive enough to measure inhibin A concentrations in the circulating blood of non-pregnant women. The study of the change of inhibin A concentrations in various physiological and pathological conditions is continuing. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (55 results)

All Other

All Publications (55 results)

  • [Publications] Y.Hasegawa: "Isolation and Characterization of Inhibin and Inhibinrelated Proteins" Journal of Reproduction and Development. 42. 23-25 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Kaneko: "Follicular Growth and FSH Secertion in Cattle" Journal of Reproduction and Development. 42. 37-40 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Mine: "Effects of Inhibin on Activin A-Induced Glucose Metabolism in Rat Hepatocytes" Endocrine. 5. 41-46 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: "Regulation of Follicle-Stimulating Hormone Receptor" Hormone Research. 46. 37-44 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Kaneko: "Inhibin is Involved in the Suppression of FSH Secretion in the Growth Phase of the Dominant-Follicle During the Early Luteal Phase in Cows" Domestic Animal Endocrinology. 14. 263-271 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.Noguchi: "Ontogeny of inhibin secretion in the rat testis:secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells" Journal of Endocrinology. 155. 27-34 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: "Dual Coupling and Down Regulation of Human FSH Receptor in CHO Cells" Life Sciences. 60. 2043-2050 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: "Expression of follicle-stimulating hormone receptor in human ovary" European Journal of Clinical Investigation. 27. 469-474 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 安部 由美子: "カレントレヴュ-1.内分泌 b.インヒビンとアクチビン" 日本産婦人科学雑誌. 48. 708-720 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 水沼 英樹: "IR-インヒビン測定キットの有用性に関する検討" ホルモンと臨床. 44. 537-545 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 長谷川 喜久: "インヒビンとアクチビンの発見とその後の展開" 獣医畜産新報. 49. 129-133 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 峯岸 敬: "FSH 受容体" 生命の科学. 48. 436-438 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 長谷川 喜久: "生殖生理を調節するインヒビンファミリー物質の役割" Hormone Frontier in Gynecology. 5. 17-26 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: "Inhibin and Activin:Novel Regulators for Gonadl Function and Sexual Maturation" Ares-Serono Symposia Publications.Rome,Itary, 16 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: "Two-Site Immunoassay for Native Inhibin A" Springer-Verlag New York,Inc., 14 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Mizunuma: "Evaluation of Maternal Serum Inhibin as a Marker for Complicated Pregnancy" Springer-Verlag New York,Inc., 11 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 長谷川 喜久: "4.3ペプチドホルモンと蛋白質ホルモンの測定法" 川島書店, 9 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: "Isolation and Characterization of Inhibin and Inhibin-related Proteins." J Reprod Dev. 42 (Suppl). 23-25 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Kaneko: "Follicular Growth and FSH Secretion in Cattle." J Reprod Dev. 42 (Suppl). 37-40 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Mine: " Effects of Inhibin on Activin A-Induced Glucose Metabolism in Rat Hepatocytes." Endocrine. 5 (1). 41-46 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: " Regulation of Follicle-Stimulating Hormone Receptor." Hormone Research. 46 (Suppl 1). 37-44 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yumiko Abe: " Inhibin and Activin." Acta Obst Gynaec Jpn. 48 (8). 708-720 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Kaneko: "Inhibin is Involved in the Suppression of FSH Secretion in the Growth Phase of the Dominant Follicle During the Early Luteal Phase in Cows." Domestic Animal Endocrinology. 14(4). 263-271 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.Noguchi: "Ontogeny of inhibin secretion in the rat testis : secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells." Journal of Endocrinology. 155. 27-34 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: "Dual Coupling and Down Regulation of Human FSH Receptor in CHO Cells." Life Sciences. 60(23). 2043-2050 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Minegishi: "Expression of follicle-stimulating hormone receptor in human ovary." European Journal of Clinical Investigation. 27. 469-474 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: Inhibin and Activin : Novel Regulators for Gonadal Function and Sexual Maturation.Ares-Serono Symposia Publications, 16 (1966)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: Two-Site Immunoassay for Native Inhibin A.Springer-Verlag New York, Inc., 14 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Mizunuma: Evaluation of Maternal Serum Inhibin as a Marker for Complicated Pregnancy. Springer-Verlag New York, Inc., 11 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Hasegawa: "Isolation and Characterization of Inhibin and Inhibin-related Proteins" Journal of Reproduction and Development. 42(Suppl). 23-25 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Kaneko: "Follicular Growth and FSII Secretion in Cattle" Journal of Reproduction and Development. 42(Suppl). 37-40 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Mine: "Effects of Inhibin on Activin A-Induced Glucose Metabolism in Rat Hepatocytes" Endocrine. 5(1). 41-46 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Minegishi: "Regulation of Follicle-Stimulating Hormone Receptor" Hormone Research. 46(Suppl 1). 37-44 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Kaneko: "Inhibin is Involved in the Suppression of FSH Secretion in the Growth Phase of the Dominant Follicle During the Early Luteal Phase in cows" Domestic Animal Endocrinology. 14(4). 263-271 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] J.Noguchi: "Ontogeny of inhibin secretion in the rat.testis:secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells" Journal of Endocrinology. 155. 27-34 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Minegishi: "Dual Coupling and Down Regulation of Human FSH Receptor in CHO Cells" Life Sciences. 60(23). 2043-2050 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Minegishi: "Expression of follicle-stimulating hormone receptor in human ovary" European Journal of Clinical Investigation. 27. 469-474 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 安部由美子: "カレントレヴュ- 1.内分泌b.インヒビンとアクチビン" 日本産科婦人科学雑誌. 48(8). 708-720 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 水沼英樹: "IR-インヒビン測定キットの有用性に関する検討" ホルモンと臨床. 44(5). 537-545 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 長谷川喜久: "インヒビンとアクチビンの発見とその後の展開" 獣医畜産新報. 49(2). 129-133 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 峯岸敬: "FSH受容体" 生命の科学. 48(5). 436-438 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 長谷川喜久: "生殖生理を調節するインヒビンファミリー物質の役割" Hormone Frontier in Gynecology. 5(1). 17-26 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Hasegawa: "Inhibin and Activin:Novel Regulators for Gonadal Function and Sexual Maturation" Ares-Serono Symposia Publications,Rome,Italy, 16 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Hasegawa: "Two-Site Immunoassay for Native Inhibin A" Springer-Verlag New York,Inc., 14 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Mizunuma: "Evaluation of Maternal Serum Inhibin as a Marker for Complicated Pregnancy" Springer-Verlag New York,Inc., 11 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Hasegawa: "Isolation and characterization of inhibin and inhibin-related proteins." J. Reprod. Develop.42(Suppl.). 23-25 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Kaneko: "Follicular growth and FSH secretion in cattle." J. Reprod. Develop.42(Suppl.). 37-40 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 長谷川喜久: "インヒビンとアクチビンの発見とその後の展開" 獣医畜産新報. 49. 129-133 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 安部由美子: "カレントレヴュ-1.内分泌b.インヒビンとアクチビン" 日産婦誌. 48(8). 708-720 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 安部由美子: "下垂体機能と成長因子" Hormone frontier in gynecology(メヂィカルビュー社). 3. 309-316 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 水沼英樹: "測定法IR-インヒビン測定キットの有用性に関する検討" ホルモンと臨床. 44(5). 537-545 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Mine: "Effects of Inhibin on Activin A-Induced Glucose Metabolism in Rat Hepatocytes" Endocrine. 5(1). 41-46 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 伊吹令人: "今日の治療 月経異常の臨床" 永井書店, 190 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 長谷川喜久: "ペプチドホルモンと蛋白質ホルモンの測定法「動物生殖機能実験の手引き」、菅原七郎監修" 川島書店, 9 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Hasegawa: "Inhibin and Activin : Novel Regulators for Gonadal Function and Sexual Maturation" Ares-Serono Symposia Publications, 16 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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