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Quatification of BMP-4 in human serum

Research Project

Project/Area Number 04670890
Research Category

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

Allocation TypeSingle-year Grants
Research Field Orthopaedic surgery
Research InstitutionOsaka University

Principal Investigator

MASUHARA Kensaku  Osaka University Medical School, Assistnat Professor, 医学部, 助手 (90238915)

Co-Investigator(Kenkyū-buntansha) ONO Keiro  Osaka University Medical School, Professor, 医学部, 教授 (70028330)
YOSHIKAWA Hideki  Osaka University Medical School, Assistnat Professor, 医学部, 講師 (60191558)
HASHIMOTO Jun  Osaka University Medical School, Assistnat Professor, 医学部, 助手 (40237938)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsMonoclonal antibody / BMP-4 / Western blot analysis / Northern blot analysis / Immunohistochemistry
Research Abstract

A monoclonal antibody which reacts with murine and human bone morphogenetic protein-4 (BMP-4) has been developed using recombinant BMP-4 as an immunogen. The antibody that bound most tightly to recombinant murine (rm) BMP-4 was selected, subcloned and characterized. The specificity of the antibody was confirmed using Western blot analysis and enzyme-linked immunosorbent assay (ELISA). The antibody reacts with murine and human BMP-4 in both the reduced and non-reduced condition, however this antibody shows cross-reactivity with neither human BMP-2 nor TGI-beta1. Thus, the produced antibody could recognized the disulfide-linked dimeric structure of bioactive BMP-4, regardless of the species. Immunocytochemical study using this antibody successfully shows the cytosolic localization of BMP-4 in osteoinductive cells, i.e., BFO and Saos-2 in which the level of mRNA for BMP-4 was proved to be constitutively high by northern blot analysis. In addition, the antibody could demonstrate the presence of BMP-4 in developmental bone formation in the alveolar bone of rat embryo by immunohistochemistry. The antibody could be used for a more sensitive approach for quantitative analysis of BMP-4.
Sandwitch ELISA for quantification of BMP-4 in sera from patients with fractures and normal controls suggessted that BMP-4 in human serum could be below 1ng/ml.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] K.Masuhara,et.all: "Use of monoclnal antibody to detect bone morphogenetic protein-4 (BMP-4)" Bone. 16. 91-96 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Masuhara,et.all: "Perioseal and intratumorous bone formation in athymic nude mice by Chinese hamster ovary tumors expressing murine bone morphogenetic protein-4." Clinical Orthopaedics and Related Research. 300. 274-280 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Masuhara,et.all: "Gene cloning and expression of a bone morphogenetic protein derived from a murine osteosarcoma (Dunn)." Clinical Orthopaedics and Related Research. 294. 344-352 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Masuhara,et.all: "Purification and characterization of bone inducing protein from a murine osteosarcoma." Clinical Orthopaedics and Related Research. 292. 329-336 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masuhara K., Nakase T., Takaoka K., Matsui M., Anderson H.C.: "Use of monoclonal antibody to detect bone morphogenetic protein-4 (BMP-4)." Bone. 16. 91-96 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shimisu K., Yoshikawa H., Matsui M., Masuhara K., Takaoka K.: "Periosteal and intratumorous bone formation in athymic nude mice by Chinese hamster ovary tumors expreesing murine bone morphogenetic protein-4." Clinical Orthopaedics and Related Research. 300. 274-280 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takaoka K., Yoshikawa H., Hashimoto J., Masuhara K., Miyamoto S., Suzuki S., Ono K., Matsui M., Oikawa S., Tsuruoka N., Tawaragi Y., Inuzuka C., Katayama T., Sugiyama M., Tsujimoto M., Nakanishi T., Nakazato H.: "Gene cloning and expression of a bone morphogenetic protein derived from a murine osteosarcoma (Dunn)." Clinical Orthopaedics and Related Research. 294. 344-352 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takaoka K., Yoshikawa H., Hashimoto J., Masuhara K., Miyamoto S., Suzuki S., Matsui M., Ono K.: "Purification and characterization of bone inducing protein from a murine osteosarcoma." Clinical Orthopaedics and Related Research. 292. 329-336 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yoshikawa H., Takaoka K., Hashimoto J., Masuhara K., Ono K.: "Diagnostic and prognostic significance of bone morphogenetic protein in osteosarcoma." In Frontiers in Osteosarcoma Research, J.F.Novak, J.H.McMaster (eds.), 504-514, Hogrefe and Huber. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Masuhara,et.all: "Use of monoclonal antibody to detect bone morphogenetic protein-4(BMP-4)" Bone. 16. 91-96 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Takaoka,H.Yoshikawa,J.Hashimoto,S.Miyamoto,K.Masuhara,H.Nakahara,M.Matsui,K.Ono: "Purification and Characterization of a Bone-Inducing Protein From a Murine Ostessarcoma(Punn Type)" Clinical Orthopaedics and Related Research. 292. 329-336 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Takaoka,H.Yoshikawa,J.Hashimoto,K.Masuhara,S.Miyamoto,S.Suzuki,K.Ono,et al: "Gene Cloning and Expression of a Bone Morphogenetic Protein Derived From a Murine Osteosarcoma." Clinical Orthopaedics and Related Research. 294. 344-352 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masuhara.K.,Suzuki.S.,Yoshikawa,H.,Takaoka,K.: "Use of monoclonal antibodies to detect wurine bone morphogenetic Protein" The 14th Annual Meeting The American Society For Bone and Mineral Research.抄録集. (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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