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Molecular Assembly and Stability of Human Hepatitis B Virus Surface Antigen Vaccine

Research Project

Project/Area Number 02453151
Research Category

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

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

Principal Investigator

TAKAGI Toshio  Osaka University. Institute for Protein Research, Professor, たんぱく質研究所, 教授 (00029943)

Co-Investigator(Kenkyū-buntansha) ISHIKAWA Nobuyoshi  Research Foundation of Microbial Diseases of Osaka University, Kanonji Institute, 微生物病研究会・観音寺研究所, 課長代理 (30113450)
KAMEYAMA Keiichi  Osaka University, Institute for Rrotein Pesearch, Instructor, たんぱく質研究所, 助手 (60177607)
佐藤 衛  大阪大学, たんぱく質研究所, 助手 (60170784)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordsvaccine / Gene Engineering / electrophoresis / Light Scattering / ヒトB型肝炎 / X線小角散乱
Research Abstract

Vaccines superior to the conventional ones can now be produced in principle using gene engineering technique. Human hepatitis B virus surface antigen vaccine derived from yeast(y-HBsAg vaccine)is, however, the first and only successful outcome of the introduction of the engineering technique into vaccine production. Its efficacy is believed to be derived from the particular nature and high stability of the assembly composed mainly of protein polypeptides and lipids. The present research project was motivated from our belief that the mode of assembly of the constituents and the source of the stability must be understood to provide basic knowledge hopefully leading to production other efficient vaccines through the approach.
The y-HBsAg vaccine particles were found to be spherical, and homogeneous with respect to their size having radius of about 19 nm as judged from dynamic light scattering, small-angle X-ray scattering and electron microscopy. No regular structure was detected in the vaccine particle by small-angle X-ray scattering. The vaccine particle was eluted from a TSK-Gel G600OPW column as a symmetrical peak. Monitoring of the elution gave molar mass estimate of 5, 040 kg/mol for the vaccine particle of the target in the present study of which constituent polypeptide has 9 additional amino acid residues in the N-terminal side compared to the polypeptide of the virus surface antigen. The molar mass of the vaccine particle consisted of the latter polypeptide was estimated to be 4, 640 kg/ mol. The vaccine particle was stable against heating up to 8O゚C. The constituent polypeptide also showed stability against heating in the presence of SDS. Isoelectric point of the particle was estimated to be 4.0 to 4.2 by electrophoretic light scattering in free solution. It is hoped that these results and approaches can play a role of guidepost for future characterization of vaccine particles expected to be developed in near future using gene engineering technique.

Report

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

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Keiichi Kameyama: "Measurement of electrophoretic mobilities of sodium dodecyl sulfate simple micelles and mixed micelles with nonionic surfactant,octaethylene glycol dodecyl ether,by electrophoretic light scattering with the correction for electroosmotic flow" J.Colloid Surface Sci.140. 517-524 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 高木 俊夫: "電気泳動光散乱法〜蛋白質から細胞までを対象とした新規装置について" 生物物理化学. 36. 1-11 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Toshio Takagi: "Characterization of yeast-derived human hepatitis B vaccine particle:Gross conformation and polypeptide composition"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takagi, T.: "Application of low-angle laser light scattering detection in the field of biochemistry" J. Chromatogr.506. 409-416 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sato, Y.: "High-performance size exclusion chromatography and molar mass measurement by low-angle laser light scattering of recombinant yeast-derived human hepatitis B virus surface antigen vaccine particles." J. Chromatogr.507. 25-31 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sato, Y.: "Characterization of yeast-derived human hepatitis B virus surface antigen vaccine particle : Gross conformation and polypeptide composition"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takagi, T.: "New aspects of electrophoresis of membrane proteins" Electrophoretic light scattering and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. 391-406 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Keiichi Kameyama: "Measurement of electrophoretic mobilities of sodium dodecyl sulfate simple micelles and mixed micelles nonionic surfactant,octaethylene glycol dodecy ether,by electrophoretic light scattering with the correction for electroosmotic flow" J.Colloid Surface Sci.140. 517-524 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] 高木 俊夫: "電気泳動光散乱法〜蛋白質から細胞までを対象とした新規装置について" 生物物理化学. 36. 1-11 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Toshio Takagi: "Characterization of yeastーderived human hepatitis B vaccine particle:Gross conformation and polypeptide composition"

    • Related Report
      1991 Annual Research Report
  • [Publications] Yoshiko Sato,Nobuyoshi Ishikawa,Mamoru Sato,Keiichi Kameyama,Toshio Takagi: "Characterization of Yeastーderived Human Hepatitis B Vaccine Particle:Gross conformation and polypeptide compositin" Journal of Biochemistry.

    • Related Report
      1990 Annual Research Report

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

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