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Clinical Chemistry 38: 2233-2238, 1992;
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Clinical Chemistry, Vol 38, 2233-2238, Copyright © 1992 by American Association for Clinical Chemistry

Sandwich immunoassay for intact human osteocalcin

K Hosoda, H Eguchi, T Nakamoto, T Kubota, H Honda, S Jindai, R Hasegawa, M Kiyoki, T Yamaji and M Shiraki
Institute for Biomedical Research, Teijin Ltd., Tokyo, Japan.

To overcome the problems of limited-region specificity associated with conventional radioimmunoassay (RIA), we developed a sandwich enzyme immunoassay (EIA) for intact human osteocalcin (hOC). For this EIA we used antibodies to the N- and C-terminal regions of hOC that were raised against an N-terminal 20-residue peptide and a C-terminal 7- residue peptide, both synthetic. Immunoassay profiles of tryptic digests of hOC and serum from patients with renal failure, fractionated by reversed-phase HPLC, facilitated direct demonstration of the region specificity of this method. A preliminary study of serum osteocalcin concentrations in patients with renal failure further confirmed this specificity, showing lower positive rates obtained by this method than by conventional RIA. The cross-reactivity data of hOC with bovine and rat osteocalcins by the sandwich method indicated its species specificity. These studies demonstrate the superior specificity of this sandwich EIA compared with conventional RIA and thus confirm its potential diagnostic superiority.


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