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Purification of Rh(D) antigen and its application for immuno-absorption therapy

Research Project

Project/Area Number 61570791
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionOsaka University

Principal Investigator

NEGORO Takao  Osaka University Medical School, 医学部, 助手 (00112053)

Co-Investigator(Kenkyū-buntansha) MASTUZAKI Noboru  Osaka University medical School, 医学部, 助手 (30199781)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsRh incompatible pregnancy / Rh(D) antigen / 抗原固定免疫吸着カラム
Research Abstract

In order to rescue the severly affected fetuses by Rh iso-immunization, we newly developed the solid phase immuno-absorbent column that eliminated effectively anti Rh(D) antibodies from immunized mothers. The results obtained in this project were the followings.
(1) The bulk of red blood cells (RBC) were obtained from Rh(-D-) patientsand stored for later use. The amount of Rh(D) antigens expressed on cell surface in Rh(-D-) patients was increased to 5 times as compared with that of normal Rh(D) person.
(2) Rh(D) antigens were purified from RBC of Rh(-D-) patients. The membrane fractions were separated by the discontinuous Sucrose gradients and solubilized by non-ionic detergent,Renex 30. The solubilized fractions were subjected to lentile lectin affinity column, followed by gel filtration in the presence of detergentt. The purification of membrane antigen was attained by 100 times at the peak fraction of antigenic activity. SDS-PAGE revealed that the solubilized zntigens were 28K-35K dalton proteins.
(3) The immuno-absorbent column was produced by the coupling of the semi-purified Rh(D) antigen to Sepharose 4B. The absorbent system thus produced was very effective because it could eliminate the anti Rh(D) antibodies from the pregnat sera possessing high titer antibodies. However, much larger amount of the purified antigens were needed for repeated clinical use of these systems. The application of the genetic engineering technique is useful to overcome these problems.

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Takao Negoro: Human Immunol.15. 137-149 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: Am.J.Reprod.Immunol.12. 13-16 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Masayasu Koyama: Acta.Obst.Gynec.Jpn.38. 2091-2092 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: Acta.Obst.Gynec.Jpn.39. 159-160 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takashi Kameda: Japan Society for Basic Reprod.Immunol.1. 56-59 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: Japan Society for Basic Reprot.Immunol.1. 73-79 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Yoshihito Samejima: Japan Society for Basic Reprod.Immunol.1. 80-86 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: Am.J.Reprod.Immunol.Microbiol.13. 121-124 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: Acta.Obst.Gynec.Jpn.39. 1655-1658 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Noboru Mastuzaki: Acta.Obst.Gynec.Jpn.40. (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Noboru Mastuzaki: Acta.Obst.Gynec.Jpn.40. (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: "Serological and functional analysis of the epitope clusters on Leu3/T4 antigens." Human Immunol.15. 137-149 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: "Further characterization of a human sperm coating antigen (gp12)." Am. J. Reprod. Immunol.12. 13-16 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Masayasu Koyama: "Low susceptibility of choriocarcinoma cell lines to lymphokine-activated killer (LAK) cells." Acta. Obst. gynec. Jpn.38. 2091-2092 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: "Heterogeneous expression of Leu3/T4 antigen on cord T cell." Acta. Obst. Gynec. Jpn.39. 159-160 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takashi Kameda: "Analysis of Leu3/T4 antigen on cord T cells." Japan Society for Basic Reproductive Immunology. 1. 56-59 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: "Lymphokine activated killer (LAK) activity against trophoblast cell lines." Japan Society for Basic Reproductive Immunol.1. 73-79 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Yoshihito Samejima: "Six monoclonal antibodies specific to trophoblast cell lines." Japan Society for Basic Reproductive Immunol.1. 80-86 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: "Effect of a soluble factor secreted from cultured trophoblast cells on in vitro lymphocyte reaction." Am.J. Reprod. Immunol. Microbiol.13. 121-124 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fumitaka Saji: "Identification of a human sperm antigen to sperm immobilizing antibodies." Acta. Obst. Gynec. Jpn.39. 1655-1658 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Noboru Mastuzaki: "The Immunosuppression of human T-cell proliferation pathways by a choriocarcinoma-cell derived factors." Acta. Obst. Gynec. Jpn.40. (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao Negoro: "Lymphokine activated killer (LAK) activity against trophoblast cell lines" Japan Society for Basic Reprod. Immuol.1. 73-79 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takao.Negoro.: Human Immunology. 15. 137-149 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Takao.Negoro.: Acta.Obst.Gynec.Jpn.39. 488-489 (1986)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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