| Project/Area Number |
60570794
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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 | Keio University |
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Principal Investigator |
MORISADA Masaru Keio University, School of Medicine, Dept. of Ob/Gyn., 医学部・産婦人科学教室, 講師 (40051552)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Kimihiko Keio University, School of Medicine, Dept. of Ob/Gyn., 医学部・産婦人科学教室, 助手 (50168340)
SUEOKA Kou Keio University, School of Medicine, Dept. of Ob/Gyn., 医学部・産婦人科学教室, 助手 (90162833)
NATORI Michiya Keio University, School of Medicine, Dept. of Ob/Gyn., 医学部・産婦人科学教室, 講師 (80101913)
TADAKUMA Takushi Keio University, School of Medicine, Dept. of Bacteriology, 医学部・微生物学教室, 助教授 (30051626)
宮崎 豊彦 慶應義塾大学, 医学部・産婦人科学教室, 助手
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1987: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1986: ¥400,000 (Direct Cost: ¥400,000)
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| Keywords | Early pregnancy factor / Purification / Column / Artificial insemination / 人工授精 / 体外受精・胚移植 / Early Pregnancy Factor / Rosette inhibition test / Chromatography |
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| Research Abstract |
Early Pregnancy Factor (EPF) is a pregnancy association factor which appears in maternal blood in the early period after fertilization. We established the EPF assay system with higher stability on a smaller scale using the 96 well microplate.
Isolation of EPF activity was carried out using crude hCG as a sourse. The activity of EPF was detected in a dose dependent mannor for biochemical analysis and considerations. Two peaks of protein were seen at 20KD and 40KD by Sephdex G-75 column. The protein that showed a peak at 40KD demonstrated both EPF and hCG activities. Using DE-52 column, EPF was isolated from its contaminated fractions of hCG. Gelpermiation chromatography and SDS-PAGE could not demonstrate the specific EPF proteinband and peak from the fraction of DE-52. Reversephase HPLC analysis showed eight peaks which included specific two peaks and the other two peaks, of which proteindose increased with EPF activity. The four peaks were determined using anamino acid analysis as an independent peptides. Fast protein liquid chromatography including anion, cation and chromatofocusing was used to isolate several EPF peaks in each of the column, the chromatofocusing showed P16,5 in the highest EPF activity fraction.
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