| Project/Area Number |
60570372
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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 |
Neurology
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| Research Institution | Kitasato University |
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Principal Investigator |
SAITO Toyokazu Kitasato University, assistant professor, 医学部, 講師 (80050574)
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| Co-Investigator(Kenkyū-buntansha) |
KANAIZUKA Fumiaki Kitasato University, Assistant, 医学部, 助手 (90169540)
MIYATA Kayoko Kitasato University, Assistant, 医学部, 助手 (40159405)
HOSODA Minoru Kitasato University, Assistant, 医学部, 助手 (30146401)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥200,000 (Direct Cost: ¥200,000)
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| Keywords | Enzyme-linked immunosorbent assay(ELISA) / CSF髄鞘蛋白(MAG / BP / Wolfgram蛋白 / proteolipid蛋白) / CSF / myelin protein |
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| Research Abstract |
The purpose of this study is to measure central nervous system(CNS)myelin protein of CSF in some neurological diseases. A sensitive and facile enzyme-linked immunosorbent assay using a microplate was developed for quantitating myelin protein and antibodie to these proteins. CNS myelin basic protein(mBP) was isolated from autopsied human brain and vobine brain and purified by the procedure of Banik and Davison(1973,J,Neurochem.). Antibodies to MBP raised in rabbits and anti-MBP antibodies were isolated by CNBr activated sepharose 4B affinity chromatography. Purified MBP or anti MBP antibodies were detected by SDS-PAGE and immunoblotting. Biotinylated anti-human IgG(1:100 dilution) and avidin-D peroxidase(1:1000) were used for ELISA. Raugh Myelin associated glycoprotein(MAG) was isolated by the method of Quarles(1982,Neurochem Research). MAG were purified by sepharose CL-6B colum chromatography. The purity of the MAG was checked by electrophoresis on SDS slab gels. Antibodied to MAG were obtained from rabbits. The rabbit IgG was isolated by DE-52 colum chromatography. Anti-MAG antibodies were isolated by phenyl -sepharose 4B affinity chromatography. Wolfgram protein of CNS myelin was also prepared from developed SDSPAGE. ANtibodies to Wolfgram protein was also prepared from rabbits. Antibodies to BmBP elevated in patients with MS and sometimes MBP was also elevated. Anti MAG antibodies was not detected in CSF in patients with demylinating disease. CSF MAG and Wolfgram protein were detected in some demylinating disese. ELISA is useful to measure the minimal values of CSF MBP,MAG or Wolfgram protein or antibodies to these protein.
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