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Purification of Common Lung Cancer Antigen recognized with Monoclonal Antibody TFS and Production of Monoclonal Antibodies of Second Generation by its antigen.

Research Project

Project/Area Number 02670338
Research Category

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

Allocation TypeSingle-year Grants
Research Field Respiratory organ internal medicine
Research InstitutionUniversity of Tokyo

Principal Investigator

FUJISAWA Michio  University of Tokyo, Health Service center, M. D., 保健センター, 助手 (60218998)

Co-Investigator(Kenkyū-buntansha) YAMANE Akira  University of Tokyo, Faculty of Medicine, M. D., 医学部第三内科, 医員
HAGIWARA Koichi  University of Tokyo, Faculty of Medicine, M. D, 医学部第三内科, 助手
OKABE Tetsuro  University of Tokyo, Faculty of Medicine, M. D, 医学部第三内科, 助手 (80169135)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsLung Cancer / Monoclonal Antiboy / Cancer Antigen / Cancer Antiyeu / Lung cancer / Monoclonal antibody
Research Abstract

Monoclonal antibody(MAb)TFS-2 was produced against Small Cell Lung Cancer and reacted with all tissue types of lung cancer. Recently we tried the radioimmunodetection of human lung cancer with ^<99m>Tc labeled TFS-2 followed by getting clear imaging of primary and metastatic lung cancers in the various tissues such as bone, lung, and liver.
(1)Purification of Common Lung Cancer Antigen.
Masses of metastatic small cell lung cancer to liver was used as source for purification. Samples were solubilized by PBS with 0.5% of NP-40. Samples were twice passed through Affinity Chromatography(tresyl-activated Sepharose 4B couppled with MAb TFS-2). NP-40 was removed by ethanol pi-ecipitatiori. SDS-PAGE showed 2 bands of protein, 39kDa and 33kDa respectedly. Band of 33 kDa reacted with MAb TFS-2 on the Western blotting. Protein of 33kDa was analyzed directly transferring to membrane and 20 amino acid residue was determined.
(2)Analysis of Gene coding Common Lung Cancer Antigen.
The MRNA was obtained from small cell lutig cancer cell line NCI-1169, and the expression CDNA library in plasmid was constructed from the MRNA. The plasmids were transfected to Cos-1 cells and clones expressing antigen on the cell surface were screened by ^<125>I labeled MAb TFS-2. A EDNA clone coding the protein reacted with MAb TFS-2 was isolated.
(3)Production of Monoclonal antibodies of the second generation Several clones of hybridoma cell that reacted with purified antigen presently were obtained.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] Yoshimi Umezawa.,et al: "Identity of Brain-Associated Small Cell Lung Cancer Antigen and the CD56(NKH/Leu-19) Leukocyte Differentiation Antigen and the Neural Cell Adhesion Molecule." Japanese Journal of Clinical Oncology. 21. 251-255 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Umezawa, S. Kuge, N, Kikyo, T. Shirai, J. Watanabe, M. Fujisawa, and T. Okabe: "Identity of Brain-associated Small Cell Lung Cancer Antigen and CD56 (NKH-1/Leu-19) leukocyte differenciation Antigen and the Neural Cell Adhesion Molecule." Jpn J Clin Oncol. 21. 251-255 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yoshimi Umezawa.,et al: "Identity of BrainーAssociated Small Cell Lung Cancer Antigen and the CD56(NKH/Leuー19)Leukocyte Differentiation Antigen and the Neural Cell Adhesion Molecule." Japanese Jounal of Clinical Oncology. 21. 251-255 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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