| Project/Area Number |
09670237
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | Keio University |
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Principal Investigator |
FUKUMA Mariko Keio University School of Medicine, Instructor, 医学部, 助手 (60101995)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Taketo Keio University School of Medicine, Instructor, 医学部, 助手 (60230463)
UMEZAWA Akihiro Keio University School of Medicine, Assistant professor, 医学部, 講師 (70213486)
HATA Jun-ichi Keio University School of Medicine, Professor, 医学部, 教授 (90051614)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | EC Cells / monoclonal antibody / glycolipid / glycoprotein / proteoglycan |
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| Research Abstract |
It has been postulated that the cell surface antigens expressed on embryonic cells play an important role in development and differentiation. The embryonic character of germ cell tumors (GCTs) provide unique opportunities to investigate embryonic cellular differentiation as well as the relation ship between developmental biology and oncology. In efforts to augment our knowledge of the differentiation mechanism involved in human GCTs several monoclonal antibodies were raised against a human embryonal carcinoma cell-line (NCR-G3) which was established from human GCT.We report here the characterization of two monoclonal antibodies 6E2-Mab and 4C4-Mab, both antibodies specifically immuno-stain embryonal carcinoma but not other types of human tumors. The antigenicity of 6E2-Ag was disappeared when it was treated with periodic acid, neuraminidase, but not with proteases or heating at 100ﾟC which suggests that 6E2-Mab recognizes carbohydrate moiety and has a sialic acid at the non-reduced terminal. 6E2-Ag was extracted into alkaline chloroform methanol mixture and was detected at the position of rf value 1.7 when it was developed on a thin layer chromatography. This means 6E2-Ag is glycolipid and 6E2-Mab recognizes carbohydrate moiety. The antigenicity of 4C4-Ag was disappeared when it was treated with proteases or periodic acid. 4C4-Ag was also disappeared from the cell surface when Tunicamycin was added to the medium. By agarose gel chromatography the molecular weight of 4C4-Ag was estimated about 1 x 1O^7kDa arid was adsorbed into DEAE-ion exchange column and eluted out with 0.5 M NaCl. When the eluate was treated with 4 M guanidine, the antigenicity peak in agarose gel chromatography shifted to the molecular weight around 2 x 10^6kDa. When the DEAE-column eluate was treated with chondroitinase ABC the antigenicity was detected at a molecular weight 85 kDa. These result suggest that 4C4-Mab recognizes the carbohydrate moiety of proteoglycan core protein.
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