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Structure of antigenic complex carbohydrate chains recongnized by carbohydrate directed monoclonal antibodies

Research Project

Project/Area Number 62580118
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionKyoto University

Principal Investigator

FUNAKOSHI Ikuo  Faculty of Pharmaceutical Sciences, Kyoto University, 薬学部, 助手 (10025702)

Co-Investigator(Kenkyū-buntansha) SUGAHARA Kazuyuki  Faculty of Pharmaceutical Sciences, Kyoto University, 薬学部, 助手 (60154449)
YAMASHINA Ikuo  Faculty of Pharmaceutical Sciences, Kyoto University, 薬学部, 名誉教授 (70025675)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1987: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsMonoclonal antibody / Carbohydrate directed monoclonal antibody / Tumor-assiciated antigen / Mucin-type glycoprotein / ムチン / 抗原糖鎖構造 / Oーグリコシド型糖鎖 / LS180 / ムチン型糖鎖 / 結腸癌細胞 / SW1118
Research Abstract

Many monoclonal antibodies that recongnize the carbohydrate moieties of glycoconjugates have been established by imumunizing mice with human colorectal cancer cells (LS180, SW1116) and assaying antibodies with glycopeptide-coated plastic assay plates. One of such antibodies, MLS102, reacted with the colorectal cancer surface and mucinous glycoproteins secreted by the cancer cells, but only weakly with normal colon tissues. By assaying the reactivities with MLS102 of submaxillary mucins and synthetic oligosaccharide glycosides, the structure of an epitopic carbohydrate recongnized by MLS102 was determined as a cluster of NeuAc 2 6GalNAc attached to a certain peptide.
MLS102 antigens were isolated from cell lysates and spent media of LS180, and were compared with MLS103 antigens which were detected on cancerous and adjacent normal colon tissues. Affinity purified MLS102 antigen contained Ser, Thr and Pro as the most abundant amino acids, and the carbohydrate chains were composed mainly of O-glycosidically linked NeuAc 2 6GalNAc (56%) and GalNAc residues (25%). O-glycosidically linked oligosaccharides of MLS103 antigen were different in size from those of MLS102 antigen, and consisted of longer carbohydrate chains.
Another monoclonal antibody MSW113 generated using SW1111 as immunogen was shown to be directed mainly to mucin type oligosaccharide with sialyl-Le^a antigen. Reactivity of MSW113 to sialyl-Le^a was stronger than that of NS19-9, which is believed to be raised against the same determinant group. MSW113 binds to sialyl-Le^a-ol, LS-tetrasaccharide a, and disialyllacto-N-tetraose with higher affinities, compared to NS 19-9. These two antibodies could clearly be distinguished in that MSW113 bound to sialic acid but not fucose, whereas NS 19-9 bound to fucose but not to sialic acid-containig internal structures. MSW113 was found to be useful for detecting antigens in the bloodstream of patients, especially those with pancreas cancer.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Akira Kurosaka: J.Biol.Chem.263. 8724-8726 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Hiroshi Kitagawa: J.Biochem.104. 591-594 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Hiroshi Kitagawa: J.Biochem.104. 817-821 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Shigeyuki Fukui: Jpn.J.Cancer Res.(Gann). 79. 1119-1129 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Akira Kurosaka: "A monoclonal antibody that recongnizes a claster of a disaccharides, NeuAc 2 6GalNAC, in mucin-type glycoproteins" J. Biol. Chem.263. 8724-8726 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Hiroshi Kitagawa: "Immunoaffinity isolation of a sialyl-Le^a oligosaccharide from human milk" J. Biochem.104. 591-594 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Hiroshi Kitagawa: "A monoclonal antibody that recongnizes sialyl-Le^a oligosaccharide, but Distinct from NS 19-9 as to epitope recongnition" J. Biochem.104. 817-821 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Shigeyuki Fukui: "Production of monoclonal antibodies directed against carbohydrate moieties of cell surface glycoproteins" Jpn. J. Cancer Res. (Gann). 79. 1119-1129 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Akira Kurosaka: J.Biol.Chem.263. 8724-8726 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Hiroshi Kitagawa: J.Biochem.104. 591-594 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Hiroshi Kitagawa: J.Biochem.104. 817-821 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Shigeyuki Fukui: Jpn.J.Cancer Res.(Gann). 79. 1119-1129 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Akira Kurosaka: FEBS Lett.215. 137-139 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Akira Kurosaka: J. Biol. Chem.

    • Related Report
      1987 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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