Purification of a new low molecular weight protein and its clinical application
| Project/Area Number |
02454499
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | JICHI MEDICAL SCHOOL |
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Principal Investigator |
KAWAI Tadashi Dept. of Medicine, Jichi Medical School, Professor, 医学部, 教授 (60048957)
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| Co-Investigator(Kenkyū-buntansha) |
OHBAYASHI T same as above, Lecturer, 医学部, 講師 (60102266)
ITOH Y same as above, Assistant Professor, 医学部, 助教授 (20129026)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | protein 1 / ELISA / low molecular weight protein / prostate / Clare cell / renal tubular dysfunction / protein 1 / 低分子蛋白 / protoin1 / Clara cell secretary protein / urine protein1 / human clara cell 10kDa Protein / 慢性腎不全 |
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| Research Abstract |
The authors have isolated a new low molecular weight protein (LMWP) (protein 1) from urine, characterized its physicochemical properties, determine it partial amino acid sequences, and thus prepared a monoclonal antibody against this protein. After establishing an enzyme-linked immunosorbent assay (ELISA) for this protein, the significance of this protein in laboratory medicine has been extensively studied.
1) Purification and characterization of a new LMWP (protein 1):An unknown protein detected on SDS-PAGE so far proved to be protein 1 after checking the reactivity of rough purified fraction with various antisera (antibody). The present project is thus modified to isolate protein 1 using the antibody as a probe. The established purification procedure comprised of ammonium sulfate precipitation, ConA affinity, gel-filtration, anti-retinol binding protein antibody coupled affinity, and an anion exchange column. Purity was confirmed by SDS-PAGE and its molecular weight was 14kDa and p1 was 4.7. Partial amino acid sequences were determined which proved to be identical with human lung Clara cell 10 kDa secretary protein.
2) Preparation of a monoclonal antibody.
3) Establishment of an ELISA:A reliable precise assay was initially developed using purified substance as standard and its monoclonal anbitody.
4) Clinical application:Protein 1 is distributed in various body fluids, in particular, serum value ranged in normal adults ranged from 2 to 20 mug/l and urine value was varied with marked sex-differences being higher in male than in female. Another extensive studies finally spot another synthetic sites as prostate. Further studies on this protein was both fundamentally and clinically in progress.
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Report
(4 results)
Research Products
(16 results)
