1989 Fiscal Year Final Research Report Summary
Purification of a novel SLE-associated human serum protein (HAKATA antigen) and preparation of a monoclonal antibody against the antigen.
| Project/Area Number |
63571079
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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 |
医学一般
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| Research Institution | Kyushu University |
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Principal Investigator |
YAE Yoshiaki Kyushu University, Faculty of Medicine, Assistant., 医学部, 助手 (80038736)
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| Co-Investigator(Kenkyū-buntansha) |
INABA Shoichi Kyushu University, Faculty of Medicine, Lecturer., 医学部, 講師 (90091305)
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| Project Period (FY) |
1988 – 1989
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| Keywords | HAKATA antigen / Human serum protein / beta2-glycoprotein / SLE |
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| Research Abstract |
In a study on precipitation reaction between human sera in agarose gel, Inaba and Okochi found that a serum originated from a patient with systemic lupus erythematosus(SLE) formed a precipitin line with a serum of other patient. The antigen defined by the antibody, however, is widely distributed in sera of healthy donors and patients, except those in which the antibody was detected. The antigen in serum as tentatively named "HAKATA" antigen. Soon it was known that the antibody against HAKATA antigen was found only in some sera of patients with SLE and that the antigen was deposited in glomerulus kidney specimens of a patient with lupus nephritis. It was also known that the amount of the antigen in sera of patients with acute hepatitis was higher than that of normal sera and it was lower in sera of patients with liver cirrhosis. We isolated several milligrams of HAKATA antigen and characterized the properties. The procedure used for the isolation were isoelectric precipitation, hydroxylapatite column, ammonium sulfate precipitation, Sephadex G-200 gel filtration, and lentil lectin affinity column chromatography. HAKATA antigen was purified 4700 times from original pooled serum. The mlecular weight determined by Sepharose 4B gel filtration was 650K daltons and subunit of 35K daltons was detected by sodium dosesyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) with beta-mercaptoethanol and numerous bands with molecular weight ranging from 35K to 650K were resolved by SDS-PAGE without beta-mercaptoethanol. S(20, water) was 12.0. E(1%, 280nm) was 11.7. The analysis of amino acid composition revealed that HAKATA antigen contains collagen-like structure in the molecule. The amount of HAKATA antigen in sera of 69 healthy laboratory personnels of this Hospital between age of 20 and 58 ranged 0.7 - 2.3 mg/dl. We obtained mouse monoclonal antibodies(IgG_1 kappa, IgG_<2a> kappa, and IgG_<2b> kappa) against HAKATA antigen.
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