| Project/Area Number |
58870081
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Mie University |
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Principal Investigator |
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| Project Period (FY) |
1983 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 1985: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1984: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1983: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | pregnancy protein / SP1 / Latex aggrutination immunoassay / LA system / FPLC system |
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| Research Abstract |
The pregnancy specific beta1-glycoprotein(SP1) was purified from the retroplacental hematoma as folloeing processures; (1)centrifuse of retroplacental hematoma and concentrating this supernatant with 40% saturated Ammonium Sulfate solution (2)Sephadex G-150 gel filtration (3)DEAE-Sephalose CL-6B column chromatography (4)Hydroxylapatite chromatography (5)SP1negative immunoadsorbent chromatography (6)mono-Q column chromatography of FPLC system. The purity of the refined SP1 with those processures was 99%. Anti-SP1-serum was obtained with immunizing to rabbits seven times.
Antibody-sensitized Latex was made with this antiserum by Eikenkagaku Co. limited and Latex aggrutination immunoassay (LA method) was attempted. The Latex particle, 0.1 <micro> m diameter, was combined with 220ng IgG fraction of antiserum per 1 <cm^2> surface area of the particle. Ammonium-buffered SP1 standard solution 100 ng/ml (Haechist A.G.) was able to detect in the settled condition; O.D.585 nm, given sample 80 <micro> l, Latex solution 300 <micro> l, reacting time 400 seconds, reacting temperature 37゜C.
The serum of early pregnant women were assayed with this method and SP1 was detected on the women of 6 weeks of gestation. 81 prepartum women were selected randomly to examine correlation between LA method and SRID method. This correlation was not good enough. This rapid assay, however, will be available clinicaly as a differencial diagnosis of ectopic pregnancy and placental function test. The higher sensitivity would be required for the mesurement of SP1 as a tumor marker on trophoblastic disease.
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