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Clinical Chemistry 40: 822-828, 1994;
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Clinical Chemistry, Vol 40, 822-828, Copyright © 1994 by American Association for Clinical Chemistry

Characteristics of a two-site immunoradiometric assay for human skeletal alkaline phosphatase in serum

K Panigrahi, PD Delmas, F Singer, W Ryan, O Reiss, R Fisher, PD Miller, I Mizrahi, C Darte and BC Kress
Department of Pathology, University of Maryland, School of Medicine, Baltimore 21201.

This two-site IRMA includes specific monoclonal antibodies for measuring skeletal alkaline phosphatase (B-ALP) in human serum. Assay calibration is based on mass units (micrograms per liter) and was established with purified B-ALP from a human osteosarcoma cell line, SAOS-2. Precision studies demonstrated intra- and interassay CVs of 3- 5% and 5-7%, respectively. Relative reactivity studies showed that the assay has a sevenfold preference for detecting B-ALP compared with the liver isoenzyme in serum. The normal reference interval for 478 healthy adults was 5-22 micrograms/L. Method comparison studies showed good correlation between this B-ALP assay (y) and commercially available electrophoretic methods (x) (y = 0.3540x + 20.5, R2 = 0.929) in a pagetic population. Temporal profiles for total ALP, this IRMA B-ALP assay, and B-ALP by electrophoresis in three pagetic patients were parallel. We conclude that this assay demonstrates good analytical performance and would be useful for the clinical assessment of metabolic bone disorders.


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