| Project/Area Number |
63570275
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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 |
Legal medicine
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| Research Institution | Kagoshima University |
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Principal Investigator |
TSUGANEZAWA Osao Kagoshima Univ., Fac. of Med., Professor, 医学部, 教授 (80064517)
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| Co-Investigator(Kenkyū-buntansha) |
AGO Kazutoshi Kagoshima Univ., Fac. of Med., Assistant, 医学部, 助手 (20102056)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1989: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | ABO Blood Group System / Human Group O Serum / Blood Group Antigen / Cross-reacting Anti-AB Antibody / Alloagglutinin / IgG / Fab / Affinity Chromatography / 合成血液型決定基糖 / O型 / 交叉反応性抗A、B抗体 / アフィニティクロマトグラフィー |
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| Research Abstract |
The isolation of the cross-reacting anti-AB antibody in human group O serum was accomplished by continuous triplicate performance of affinity chromatography on alternate group A and group B active columns. The preparation evenly agglutinated A and B erythrocytes and did not exhibit titer reduction by treatment with 2-mercaptoethanol. The preparation migrated as a single band on polyacrylamide electrophoresis and formed a single band in agar gel immunodiffusion against anti-human IgG antiserum. Thus, a yield of 13 mg of purified cross-reacting anti-AB antibody was obtained from 3600 ml of pooled human O sera.
Inhibition of hemagglutination of A and B erythrocytes by the antibody with A and B secretor saliva, A and B group specific substances, and A and B synthetic group specific trisaccharide-albumin conjugates showed that every one of these group specific materials inhibited the agglutination of homologous erythrocytes only. The antibody was completely adsorbed on both an affinity support containing the synthetic group A trisaccharide and that containing the B trisaccharide. Eluates from the affinity supports were capable of causing agglutination of A, B and AB erythrocytes and failed to agglutinate O erythrocytes. Fab fragments prepared from the antibody by papain digestion were almost entirely adsorbed on both group A and group B active columns. Further, indirect hemagglutination by use of anti-Fab antiserum revealed that the Fab fragments eluted from the above columns were able to combine with A, B and AB erythrocytes and failed to combine with O erythrocytes.
In conclusion, the present results suggest that (1) the so-called cross-reacting anti-AB antibody is a polyfunctional or polyspecific antibody, not a cross-reacting antibody; and (2) the antibody in question is IgG antibody.
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