Preissner, Carol M.

Abstract In this sequential assay, the thyroglobulin in serum binds to polystyrene beads coated with two mouse monoclonal antibodies. These beads then react with a rabbit polyclonal antibody, biotinylated sheep anti-rabbit IgG, streptavidin-horseradish peroxidase, and a peroxidase substrate to yield a colored product that is measured spectrophotometrically at 492 nm. The range of the standard curve is 1.6 to 100 micrograms/L. The detection limit of the assay is 1.2 micrograms/L. The interassay coefficient of variation is 14.0% at 6.2 micrograms/L, 5.7% at 32.7 micrograms/L; the intra-assay CVs range from 7.8% to 14.8%. The reference intervals are 2.7 to 42.1 micrograms/L for euthyroid persons and less than or equal to 5 micrograms/L for athyreotic patients not on thyroxin replacement therapy. Some autoantibodies to thyroglobulin cause thyroglobulin values to be falsely low. The concentration of autoantibodies is not correlated with the analytical recovery of human thyroglobulin. The coefficient of correlation between the measurement of thyroglobulin by this assay and by an immunoradiometric assay was 0.969 for 186 autoantibody-negative samples, 0.963 for 37 autoantibody-positive samples.

Abstract This new bioassay for parathyrin (PTH) in plasma (bio-PTH) combines immunoextraction on affinity columns [goat anti-hPTH (1-44) conjugated to Sepharose 4B] and a receptor assay involving an osteosarcoma cell line. The mean extraction efficacy ranges from 87% (as determined with immunopurified 125I-labeled PTH) to 62% for hPTH bioactivity. The assay is standardized with synthetic hPTH (1-84) and can detect as little as 0.9 pmol/L of PTH in 2 mL of plasma. In 100 healthy adults, the 95% reference interval for bio-PTH was less than 0.9 to 6.1 pmol/L (median, 2.0 pmol/L). In 185 patients with surgically confirmed hyperparathyroidism, bio-PTH concentrations ranged from 1.0 to greater than 120 pmol/L (median, 12.9 pmol/L); 80% of values were greater than 6.1 pmol/L. In 50 patients with both preoperative and postoperative determinations, the mean (+/--SD) concentrations of calcium in serum were 113 +/- 10 and 89 +/- 6 mg/L, respectively; the median bio-PTH concentrations were 13.6 and 2.0 pmol/L, respectively. In 22 patients with nonparathyroid-mediated hypercalcemia, the concentration of bio-PTH ranged from less than 0.9 to 5.3 pmol/L (median, 1.8 pmol/L). This bio-PTH assay is slightly less sensitive than our GP235 immunoreactive PTH (iPTH) immunoassay for detecting hyperparathyroidism (Clin Chem 1982;28:69-74); however, the bioassay is more specific and detected some cases missed by the iPTH assay. Overall, 95% of the hyperparathyroid patients had an increased test result for either the bio-PTH or the iPTH assay.

Abstract The immunochemiluminometric assay described uses immobilized anti-human parathyrin (parathyroid hormone, hPTH)(1-44) and anti-hPTH(44-68) antisera and acridinium ester-labeled anti-hPTH(1-34) to simultaneously measure both intact hPTH and its amino-terminal fragments. Results by the assay correlate well with those by a cAMP-based bioassay and the Nichols Allegro immunoradiometric assay. The minimal detection limit is 0.08 pmol/L. The normal range is 1.0-5.0 pmol/L, and values are higher in older women. About 90% of study patients with surgically proven parathyroid adenomas had above-normal preoperative PTH concentrations, whereas patients with hypercalcemia of malignancy had normal or suppressed values. This assay was designed to detect both intact PTH and amino-terminal PTH fragments; however, chromatographic fractionation of pools of primary and secondary hyperparathyroid plasma showed virtually no amino-terminal fragment activity. Nonetheless, the design is important because the absence of carboxyl-terminal binding sites prevents interference by carboxyl-terminal fragments and because bioactive amino-terminal fragments will react in the assay if they are present in the patients' sera or if they are produced by in vitro proteolysis of intact PTH.