| Project/Area Number |
01870025
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Yamaguchi University |
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Principal Investigator |
TAKEO Kazusuke Yamaguchi University School of Medicine, Professor, 医学部, 教授 (10034901)
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| Co-Investigator(Kenkyū-buntansha) |
MIMURA Yusuke Yamaguchi University School of Medicine, Assistant, 医学部, 助手 (00219718)
TANAKA Tatehiko Yamaguchi University School of Medicine, Assistant, 医学部, 助手 (20144925)
NAKAMURA Kazuyuki Yamaguchi University School of Medicine, Associate Professor, 医学部, 助教授 (90107748)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1990: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1989: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Affinity electrophoresis / Separation of anti-hapten antibodies / Thermostatic electro-blotting apparatus / Two-dimensional electrophoresis. / 03thermostatic capillary isoelectric focusing apparatus / 04thermostatic semi-micro-slab gel electrophoresis apparatus / 抗ハプテン抗体の多様性 / 抗ハプテン抗体の交差反応 / オリゴ・クロ-ナル抗ハプテン抗体 / 2次元親和電気泳動法 / 抗DNP抗体 / 抗体の多様性 / 抗体の完全分離 |
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| Research Abstract |
A high resolution two-dimensional affinity electrophoresis has been developed, using capillary isoelectric focusing as the first electrophoresis and slab gel affinity electrophoresis as the second electrophoresis. By this method anti-dinitropheny antibodies have been separated completely into several hundred homogeneous immunoglobulin G (IgG) spots. They are grouped into a number of families which are composed of several IgG spots having the same affinity to the hapten but a different isoelectric point. It is suggested that each individual family is derived from one monoclonal antibody producing cell line.
The molecular heterogeneity of rabbit anti-hapten antibodies has been investigated by this two-dimensional affinity electrophoresis apparatus. Anti-dansyl and anti-arsanilic diago-antibodies were separated also into several hundred IgG spots as in the case of anti-DNP antibodies. They were grouped into a number of monoclinal IgG families.
The two-dimensional affinity electrophoresis apparatus presented here offers a powerful tool for the elucidation of fundamental problems of immunochemistry, such as antibody heterogeneity, their production mechanism and antigen-denpendent somatic mutation and immunotolerance.
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