Measurement of a more stable region of osteocalcin in serum by ELISA with two monoclonal antibodies
作者： C Rosenquist , P Qvist , N Bjarnason , C Christiansen
DOI: 10.1093/CLINCHEM/41.10.1439
关键词: Endocrinology 、 Serum concentration 、 Chemistry 、 Internal medicine 、 Monoclonal antibody 、 Osteocalcin 、 Microgram 、 Postmenopausal women 、 Amino acid 、 Capture antibody 、 Serum samples
摘要: Intact human osteocalcin purified from femoral bones as well tryptic fragments of the intact molecule [amino acids (aa) 1-19, 20-43, and 45-49] were used to raise screen monoclonal antibodies (MAbs). A two-site ELISA for measurement in serum was developed with use these MAbs. MAb recognizing midregion (aa20-43) capture antibody, an NH2 terminus (aa7-19)-specific peroxidase-conjugated detection. Human obtained bone calibration, parallelism observed samples, NH2-terminal midfragments (aa1-43), synthetic osteocalcin. Both inter- intraassay variations < 7%. Serum healthy premenopausal women (n = 49) 18.3 +/- 4.2 micrograms/L (mean SD) 28.6 9.7 early postmenopausal 114). The mean concentration 10) decreased by 10% after 7 days storage at 4 degrees C, whereas reduced 63%. N-MID IRMA measuring monitor effect hormone replacement therapy a retrospective study. significant decrease detected only ELISA.
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