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SEA-scFv as bifunctional antibody : Construction of the bacterial expresion system and its function

Research Project

Project/Area Number 09671277
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Digestive surgery
Research InstitutionTohoku University

Principal Investigator

SUZUKI Masanori  Tohoku Univ.hospital, Research Associate, 医学部・附属病院, 助手 (70206530)

Co-Investigator(Kenkyū-buntansha) ENDO Kojin  Tohoku Univ.hospital, Research Associate, 医学部・附属病院, 助手 (70292315)
UNNO Michiaki  Tohoku Univ.School of Med., Research Associate, 医学部, 助手 (70282043)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsBiPe duct cancer / Adoptive immunotherapy / MUC-1 / MUSE II / SEA / scFu / SEA D227 / MUSE 11 / CTL / LAK / MUC-1 / バイスペシフィック抗体
Research Abstract

A SEA-antibody single chain Fv (SEA-scFv) fusion protein was produced byu bacterial expression system in this study. SEA-scFv has both staphylococcal enterotoxin A(SEA) effects and antibody activity directed at the epithelial mucin core protein MUCl, a cancer associated antigen. It was expressed mostly in the cytoplasm as an insoluble form. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using metal-chelating chromatography. The resulting SEA-scFv fusion protein preparation was found to react with MUC1 and MHC class II antigens and had the ability to enhance cytotoxicity of lymphokine activated killer cells with a T cell phenotype against a human bile duct carcinoma cell line, TFK- 1, expressing MUC1. This genetically engineered SEA-scFv fusion protein promises to be an important reagent for cancer immunotherapy.

Report

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

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Katayose Y,et al.: "Promoting Apoptosis : A novel activity associated with the cyclin-dependent kinase inhibitor p27." Cancer Research.57. 5441-5445 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shinoda M,et al.: "Effective adoptive immunotherapy by T-LAK cells retargeted with bacterial superantigen-conjugated antibody to MUCI in enografted severe combinded" Cancer Research.58. 2838-2843 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tominaga T,et al.: "Establishment of an activated macrophage cell line, A-THP-1, and its properties." Tohoku J.Exp.Med.186. 99-119 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai N,et al.: "SEA-scFv as bifunctional antibody" Construction of the bacterial expression system and its functional analysis." Biochem Biophys Res Commun. 256. 223-230 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katayose Y,Kim M,Amol N,Rakkar S,Zhuangwu Li, Kenneth H.Cowan, Prem Seth: "Promoting Apoptosis : A novel activity associated with the cyclin-dependent kinase inhibitor p27." Cancer Research. 57. 5441-5445 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shinoda M,Kudo T,Suzuki M,Katayose Y,Sakurai N,Saeki H,Kodama H,Fukuhara K,Imai K,Hinoda Y,Matsuno S: "Effective adoptive immunotherapy by T-LAK cells retargeted with bacterial superantigen-conjugated antibody to MUCI in xenografted severe combinded" Cancer Research. 58. 2838-2843 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tominaga T,Suzuki M,Saeki H,Matsuno S,Tachibana T,Kudo T: "Establishment of an activated macrophage cell line, A-THP-1, and its properties" Tohoku J.Exp.Med.186. 99-119 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai N,Kudo T,Suzuki M,Tsumoto K,Takemura S,Kodama H,Ebara S,Teramae A,Katayose Y,Shinoda M,Matsuno S,Kumagai I: "SEA-scFv as bifunctional antibody "Construction of the bacterial expression system and its functional" analysis" Biochem Biophys Res Commun. 256. 223-230 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masao Shinoda: "Effective adoptive immunotherapy by T-LAK Cells Retargeted with Bacterial Superantigen-conjugated antibody to MCC1 in Xenografiec severe combined immunodeficient mice" cancer research. 58. 2838-2843 (1998)

    • Related Report
      1998 Annual Research Report

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

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