Radioimmunoassay of human parathyroid hormone in serum

Arnaud, Claude D.; Tsao, Hang S.; Littledike, Travis

doi:10.1172/jci106476

Cited by 600 publications

(180 citation statements)

References 21 publications

(16 reference statements)

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“…Solid line indicates the regression line and dashed lines represent 95% confidence intervals on the observation for the value from patients with primary hyperparathyroidism (shown by closed triangles). The relationship between 24-h urinary cAMP and serum PTH (assayed with antiserum CH 14M [10]) was highly significant (r = 0.86, P < 0.001).…”

Section: Studies Under Synthetic Dietmentioning

confidence: 94%

The Hypercalciurias CAUSES, PARATHYROID FUNCTIONS, AND DIAGNOSTIC CRITERIA

Pak¹,

Oata²,

Lawrence³

et al. 1974

J. Clin. Invest.

497

137

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A B S T R A C T The causes for the hypercalciuria and diagnostic criteria for the various forms of hypercalciuria were sought in 56 patients with hypercalcemia or nephrolithiasis (Ca stones), by a careful assessment of parathyroid function and calcium metabolism. A study protocol for the evaluation of hypercalciuria, based on a constant liquid synthetic diet, was developed. In 26 cases of primary hyperparathyroidism, characteristic features were: hypercalcemia, high urinary cyclic AMP (cAMP, 8.58+3.63 SD gmol/g creatinine; normal, 4.02±0.70 umol/g creatinine), high immunoreactive serum parathyroid hormone (PTH), hypercalciuria, the urinary Ca exceeding absorbed Ca from intestinal tract (CaA), high fasting urinary Ca (0.2 mg/mg creatinine or greater), and low bone density by 'I photon absorption. The results suggest that hypercalciuria is partly secondary to an excessive skeletal resorption (resorptive hypercalciuria). The 22 cases with renal stones had normocalcemia, hypercalciuria, intestinal hyperabsorption of calcium, normal or low serum PTH and urinary cAMP, normal fasting urinary Ca, and normal bone density. Since their CaA exceeded urinary Ca, the hypercalciuria probably resulted from an intestinal hyperabsorption of Ca (absorptive hypercalciuria). The primacy of intestinal Ca hyperabsorption was confirmed by responses to Ca load and deprivation under a metabolic dietary regimen. During a Ca load of 1,700 mg/day, there was an exaggerated increase in the renal excretion.of Ca and a suppression of cAMP excretion. The urinary Ca of 453±+154 SD mg/day was significantly higher than the control group's 211+42 mg/ day. The urinary cAMP of 2.26±0.56 umol/g creatinine was significantly lower than in the control group. In contrast, when the intestinal absorption of calcium was limited by cellulose phosphate, the hvpercalciuria was kc'ceiz(l for plblicatifn 9 July 1073 and in rezxsed form 11 September 1973. corrected and the suppressed renal excretion of cAMP returned towards normal. Two cases with renal stones had normocalcemia, hypercalciuria, and high urinary cAMP or serum PTH. Since CaA was less than urinary Ca, the hypercalciuria may have been secondary to an impaired renal tubular reabsorption of Ca (renal hypercalciuria). Six cases with renal stones had normal values of serum Ca, urinary Ca, urinary cAMP, and serum PTH (normocalciuric nephrolithiasis). Their CaA exceeded urinal Ca, and fasting urinary Ca and bone density were normal. The results support the proposed mechanisms for the hypercalciuria and provide reliable diagnostic criteria for the various forms of hypercalciuria.

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Section: Studies Under Synthetic Dietmentioning

confidence: 94%

The Hypercalciurias CAUSES, PARATHYROID FUNCTIONS, AND DIAGNOSTIC CRITERIA

Pak¹,

Oata²,

Lawrence³

et al. 1974

J. Clin. Invest.

497

137

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“…Analytical disc-gel electrophoresis was lerformed in 8 l\i urea at pH 4.4 (19). Immunoassays were l)erformed by the procedure of Arnaud et al (20).…”

Section: Methodsmentioning

confidence: 99%

“…In amino-terminal fragments from inactive fragments has so far been impossible. The second difficulty has been the utilization of heterolcgous assays that use radioactively labeled bovine hormone as the tracer, and antibodies lprepared against the bovine or porcine hormone (28)(29)(30)20). The sensitivities of these assays are variable, and depend on the cross reactivity of the particular antiserum with the human hormone.…”

Section: Methodsmentioning

confidence: 99%

“…Analytical disc-gel electrophoresis was lerformed in 8 l\i urea at pH 4.4 (19). Immunoassays were l)erformed by the procedure of Arnaud et al (20).Automated Edman degradations were )erformed with the Beckman Sequencer, model 890B, in 1 MI Quadrol buffer.The phenylthiohydantoin (PTH) amino acids were identified by regeneration to the constituent amino acid by hydrolysis with hydroiodic acid for 20 hr at 130°(21), gas-liquid 3585 …”

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confidence: 99%

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Human Parathyroid Hormone: Amino-Acid Sequence of the Amino-Terminal Residues 1-34

Brewer

Fairwell

Ronan

et al. 1972

Proc. Natl. Acad. Sci. U.S.A.

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Human parathyroid hormone has been isolated in highly purified form from human parathyroid adenomas. The primary sequence of the amino-terminal 34 residues of the human hormone was obtained by automated degradation with a Beckman Sequencer. The phenylthiohydantoin amino acids were identified by gas chromatography and mass spectrometry. The first 34 residues of human parathyroid hormone differ from the bovine hormone by six residues, and from the porcine hormone by five residues. The amino-terminal residue is serine, similar to the porcine parathyroid hormone; bovine parathyroid hormone contains an amino-terminal alanine. Human parathyroid hormone contains two methionine residues, similar to the bovine species, whereas porcine parathyroid hormone contains a single methionine residue. Amino-acid residues in the first 34 that are unique to the human sequence include an asparagine at position 16, glutamine at position 22, lysine at position 28, and a leucine at position 30.During the last few years, a significant core of information has been obtained by several laboratories on the chemistry, biosynthesis, and secretion of parathyroid hormone (PTH). These studies have indicated that this hormone is initially synthesized as a prohormone, proparathyroid hormone (1-4). Proparathyroid hormone contains about 106 amino acids, and has an apparent molecular weight of 12,500 (4). The prohormone is rapidly converted into the storage or glandular form of the hormone, which consists of 84 amino acids and has a molecular weight of 9500. The complete amino-acid sequences of the 84 amino-acid parathyroid hormone from bovine (5, 6) and porcine (7) species have been reported. After appropriate physiological stimuli, the 9500 molecular weight form of the parathyroid hormone is secreted into the circulation (8). Shortly after entering the peripheral circulation, the glandular form of the hormone is cleaved into smaller fragments. Gel filtration of human hyperparathyroid serum by several investigators has revealed a major immunoreactive fragment(s), with a molecular weight of 5000-8000, and several minor components (8-10). Immunochemical heterogeneity of the circulating human parathyroid hormone, presumably due to the different molecular forms of PTH, was initially reported by Berson and Yalow (11), and has been confirmed by others (12, 13). The specific site(s) of cleavage in the 84-amino-acid polypeptide chain of the parathyroid hormone in the general circulation is unknown. A biologiAbbreviation: PTH, parathyroid hormone cally active pel)tide fragment of bovine 1PTH, prepared by dilute acid cleavage, has been relported (14, 15), a result indicating that the intact 84-amino-acid polyl)eptide is not needed for biological activity. This peptide has been identified as the amino-terminal peptide of the hormone, and is composed of the initial 30 residues of the sequence (15). Synthetic lel)tides of the first 34 residues of the bovine hormone (16) and the initial 30 residues of the porcine hormone (17) have been prepared and are bio...

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“…Since Berson et al reported the radioimmunoassay of parathyroid hormone (PTH) in human plasma and tissue (Berson et al, 1963), there have appeared several papers concerning the human PTH radioimmunoassay (Reiss and Canterbury, 1968;Schopman et al, 1970;Arnaud et al, 1971;Segre et al, 1972;Hori et al, 1972;Reity and Weinstein, 1973). However, human PTH is hard to obtain even in small quantity to use for the iodination.…”

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confidence: 99%

Radioimmunoassay of Parathyroid Hormone in Human Plasma and Tissue Using Commercially Available Antiserum and Modified Iodination

et al. 1974

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SynopsisThe human PTH radioimmunoassay has been developed by use of bovine PTH and anti-bovine PTH antiserum, both of which were commercially available. The iodination process was modified as to use acid solution resulting in the more stable 125I-PTH. However, the PTH radioimmunoassay employed this antiserum with 125I-bovine PTH revealed that the N-terminal portion of PTH molecule was reactive only slightly in this system, indicating that most of PTH determined by this radioimmunoassay might be an biologically inactive PTH.Plasma from normal subjects and patients with primary hyperparathyroidism or uremia showed dilutional curves parallel to the curve of the standard bovine PTH, indicating the immunological identity between human and bovine PTH in this radioimmunoassay system. The extract from the parathyroid adenoma showed an identical dilutional curve, but the extract from the ectopic PTH producing tumor showed a different dilutional curve from that of the standard PTH, which might indicate the immunochemical heterogeneity of PTH produced from the ectopic source. Plasma PTH levels obtained from normal subjects and patients with primary hyperparathyroidism, uremia or pseudohypoparathyroidism were compatible with those reported previously. These data indicate that the human PTH radioimmunoassay can be done by use of commercially available anti-bovine PTH antiserum and bovine PTH in conjunction with the modified iodination process and double antibody method. However, this assay was directed mainly to the C-terminal portion of PTH molecule and the PTH radioimmunoassay specific for the N-terminal portion will be required to evaluate the PTH regulation more precisely in man.

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Radioimmunoassay of human parathyroid hormone in serum

Cited by 600 publications

References 21 publications

The Hypercalciurias CAUSES, PARATHYROID FUNCTIONS, AND DIAGNOSTIC CRITERIA

The Hypercalciurias CAUSES, PARATHYROID FUNCTIONS, AND DIAGNOSTIC CRITERIA

Human Parathyroid Hormone: Amino-Acid Sequence of the Amino-Terminal Residues 1-34

Radioimmunoassay of Parathyroid Hormone in Human Plasma and Tissue Using Commercially Available Antiserum and Modified Iodination

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