| Project/Area Number |
61570791
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Osaka University |
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Principal Investigator |
NEGORO Takao Osaka University Medical School, 医学部, 助手 (00112053)
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| Co-Investigator(Kenkyū-buntansha) |
MASTUZAKI Noboru Osaka University medical School, 医学部, 助手 (30199781)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| 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)
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| Keywords | Rh incompatible pregnancy / Rh(D) antigen / 抗原固定免疫吸着カラム |
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| 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.
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