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Characterization of embryonic carcinoma-specific antigen 6E2

Research Project

Project/Area Number 09670237
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Experimental pathology
Research InstitutionKeio University

Principal Investigator

FUKUMA Mariko  Keio University School of Medicine, Instructor, 医学部, 助手 (60101995)

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)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
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)
KeywordsEC Cells / monoclonal antibody / glycolipid / glycoprotein / proteoglycan
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.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report

URL: 

Published: 1997-04-01   Modified: 2016-04-21  

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