Sandwich immunoassay specific for the N-terminal sequence of osteocalcin

Hiroshi Eguchi, Kenji Hosoda, Noriyoshi Kurihara, Tadakatu Nakamoto, Shinji Kobayashi, Mitchitsune Arita, Ryouichi Hasegawa, Tomohiro Ohta, Mamoru Kiyoki, Teizo Yamaji, Masataka Shiraki

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13 Scopus citations

Abstract

A monoclonal antibody (10B) against the N-terminal sequence of human osteocalcin was selected to characterize its epitope and species specificity. The cross-reactivity of 10B with human and rat osteocalcin demonstrated that the reactivities of 10B with both human and rat osteocalcins were very similar. The pin technology method was used to determine the epitope and clearly demonstrated that the epitope recognized by 10B was localized to residues 12-16, the sequence of which is identical in rat and human osteocalcin molecules. This monoclonal antibody was found to be useful for designing region-specific sandwich immunoassay systems for human and rat N-terminal osteocalcin (N-OC) molecules using rabbit anti-hN20 and anti-rN20 polyclonal antibodies, respectively. The osteocalcin levels in serum determined by this N-OC method were stable during prolonged storage at 25°C and the method could be usefully applied in the development of immunoassay systems for many osteocalcin molecules from many other species.

Original languageEnglish (US)
Pages (from-to)231-240
Number of pages10
JournalJournal of Immunological Methods
Volume184
Issue number2
DOIs
StatePublished - Aug 18 1995

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Keywords

  • N-terminal osteocalcin
  • Sandwich immunoassay
  • Synthetic peptide

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Eguchi, H., Hosoda, K., Kurihara, N., Nakamoto, T., Kobayashi, S., Arita, M., Hasegawa, R., Ohta, T., Kiyoki, M., Yamaji, T., & Shiraki, M. (1995). Sandwich immunoassay specific for the N-terminal sequence of osteocalcin. Journal of Immunological Methods, 184(2), 231-240. https://doi.org/10.1016/0022-1759(95)00094-Q