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Circulating Intact Parathyroid Hormone Measured by a Two-Site Immunochemiluminometric Assay^*

Department of Medical Biochemistry, University of Wales College of Medicine Cardiff, CF4 4XN United Kingdom

Address all correspondence and requests for reprints to: Dr. Richard C. Brown, Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, CF4 4XN United Kingdom.

A direct immunoassay for circulating intact human PTH (hPTH) is described. The method relies on the formation of an immune complex of labeled antiamino-terminal PTH antibody, intact hPTH, and solid phase antimidregion PTH antibody. A chemiluminescent aryl acridinium ester is used as label. Serum samples (100 µL) are incubated with labeled antibody, and subsequently the bound fraction is separated by the addition of solid phase antibody. The bound luminescence s i quantitated in an automatic luminometer. Luminescence intensity is directly proportional to the amount of intact PTH present in the sample. Only intact PTH was found to react in this system; there was no significant interference from PTH fragments. The assay detection limit of 0.8 pmol/L hPTH-(1–84) allowed detection of intact PTH in the serum of all normal subjects tested. A clear distinction was found between hypercal-cemic individuals subsequently proven to have primary hyper-parathyroidism and those with malignancies. The assay offers several advantages over previously described PTH immunoas-says with regard to specificity, rapidity, and reagent stability. It, thus, provides a valuable means of investigating parathyroid physiology and clinical disorders of extracellular calcium metabolism.

^* This work was supported by the Medical Research Council, the Welsh Office, and Bioanalysis Ltd.

Received September 30, 1986.

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