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Development of recombinant high affinity anti-cytokine antibody

Research Project

Project/Area Number 07557375
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section試験
Research Field 医薬分子機能学
Research InstitutionInstitute of Clinical Medicine, University of Tsukuba

Principal Investigator

SUZUKI Hiroshi  Inst.of Clinical Medicine, Univ.of Tsukuba, Lecturer, 臨床医学系, 講師 (00179243)

Co-Investigator(Kenkyū-buntansha) TSUCHIYA Masayuki  Chugai pharmacentical Co., Senior Invpstigator, 探索研究所, 研究主査
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
KeywordsPhage-display method / recombinant antibody / Anti-cytokine therapy / anti-IL-1alpha antibody / ファージディスプレイ法 / サイトカイン / 抗体工学 / 自己抗体
Research Abstract

By a phage-display method, antibody (Fab fragment) expressing phage libraries were constructed from peripheral blood lymphocytes of a patient having high titer of anti-IL-1alpha IgG autoantibodies in her serum. By 4 cycles of panning, anti-IL-1alpha Fab expressing phages were isolated and soluble Fab fragments produced from cloned Fab-phages were examined for their binding to IL-1alpha. Of over 50 Fab clones, only several clones revealed significant binding to IL-1alpha. From these findings, we postulated that misfolding during synthesis of Fab molecules is the reason why low incidence of functional Fab molecules. To examine this possibility, we produced soluble Fab molecules from cloned Fab-phages derived from monoclonal anti-DNA antibody (IgM) producing cell line (NE-1) and examined the bindings of individual Fab clones to DNA.Again, only several Fab clones of over 50 clones revealed weak bindings to DNA.Although soluble Fab were less functional, Fab fragments expressed on phages constantly bound to DNA at very low concentrations (-100ng/ml). Accordingly, Fab molecules expressed on the surface of phages kept functions of antibody activities, whereas soluble Fab molecules were much less functional. From these findings, we consider that the affinity of our recombinant anti-IL-1alpha antibodies (Fab) made by this method may be much higher than the affinity evaluated previously and so we are reevaluating the recombinant anti-IL-1alpha Fab fragments made by this method previously with Fab-phages, but not with soluble Fab molecules. If affinity of the recombinant anti-IL-1alpha Fab expressed on phages is very high, we intend to produce recombinant anti-IL-1alpha IgG molecules by the conventional method for making chimera IgG molecules.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Suzuki H.et al.: "Molecular cloning at anti-ss-A/Ro 60-kd pertide Fab fragments from infiltrating salivary gland lymphocytes of a patient with sjogren's syndrome" Biochem.Biophys.Res.Commun.(印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Suzuki H.et al.: "Interlenkin-1 receptor antagonist gene polymorphism in Japanese patients with systemic luphs erythematosus" Arthritis Rheum.40. 389-390 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Suzuki H., et al.: "Molecular cloning of anti-SSA/RO 60-kd peptide Fab fragments from infiltrating salvary gland lymphocytes of a patient with sjogren's syndrome." Biochem.Biophys.Res.Commun.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Suzuki H.et al.: "Interlenkin-1 receptor antagonist gene polymorphism in Japanese patients with systemic Inpus erythematosus." Artbritis Rheum.40. 389-390 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hiroyuki Takemura: "Ehhanced response to platelet-derived growth faitor (PDGF) in IL-6 production by cultured fibrohlasts from patients with systemil scleresis" Artbritis and Rheumatism. 39・9(Suppl). S234- (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroshi Suzuki: "Charactenization of recomninant anti-SS-A autcantibcdies derived from infiltrating lymphocytes in salivary glands of sjogrens syndrome." Artbritis and Rheumatism. 39・9号(Suppl). S287- (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroshi Suzuki: "Interleukin-1 receptor antagcnist gene pelymorphism and systemic lupus erytnematosus in Japanese patients" Artbritis and Rheumatism. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroshi Suzuki: "Molecular cloning 4 anti-SS-A/Ro 60-Kd peptide Fab fragments from infiltrating salivary gland lymphocytes of a patient with sjogren's syndrome" Bichem.Biophys.Res.Commun.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 鈴木博史: "IL- 1receptor antogonist in active systemic lupus erythematosus. A good corvelation of serum IL- 1rd concentrotion with disease aitivity and enhanced production" Arthritis and Rheumatison. 38. 1055-1059 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 鈴木博史: "IL- 1レセプターアンタゴンスト" 臨床免疫. 27. 141-146 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 鈴木博史: "サイトカインアンタゴンスト" 臨床免疫. 27. 1094-1098 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 鈴木博史: "IL- 1レセプターアンタゴニスト" 炎症と免疫. 3. 571-576 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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