Development and validation of a novel cell-based assay for potency determination of human parathyroid hormone (PTH)

Hohenstein, Axel; Hebell, Meike; Zikry, Heidi; Ghazaly, Maria El; Müeller, Frank; Rohde, Jan

doi:10.1016/j.jpba.2014.06.004

Cited by 9 publications

(12 citation statements)

References 21 publications

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“…A 2x20 kDa mPEG moiety was coupled to the N‐terminus of PTH(1‐34) via a nonhydrolyzable 6‐mercaptohexanoic acid linker to investigate the level of PTH 1 receptor (PTH1R) activity after PEGylation. The residual bioactivity of this permanently PEGylated PTH(1‐34) conjugate was determined in HEK293 cells overexpressing PTH1R . The activity of TransCon PTH was low, with essentially no biological potency compared with a PTH(1‐34) reference sample.…”

Section: Resultsmentioning

confidence: 99%

“…04/200). The assay is a cell‐based potency assay for PTH concentration‐dependent cyclic adenosine monophosphate (cAMP) production in HEK293 cells overexpressing PTH1R as described by Hohenstein and colleagues (Bioassay ‐ Labor für Biologische Analytik GmbH, Heidelberg, Germany) . The activity of released PTH(1‐34) was determined after incubation of TransCon PTH in aqueous buffer at pH 10.1 and 5°C for 3 days.…”

Section: Methodsmentioning

confidence: 99%

“…The assay is a cell-based potency assay for PTH concentration-dependent cyclic adenosine monophosphate (cAMP) production in HEK293 cells overexpressing PTH1R as described by Hohenstein and colleagues (Bioassay -Labor für Biologische Analytik GmbH, Heidelberg, Germany). (12) The activity of released PTH(1-34) was determined after incubation of TransCon PTH in aqueous buffer at pH 10.1 and 5°C for 3 days. The sample solution containing released PTH(1-34) was acidified and the concentration of PTH(1-34) was determined by ultra-highperformance liquid chromatography (UHPLC) with detection at 215 nm and tested in the biopotency assay.…”

Section: Bioactivity Assaymentioning

confidence: 99%

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Design and Preclinical Development of TransCon PTH, an Investigational Sustained-Release PTH Replacement Therapy for Hypoparathyroidism

Holten-Andersen

Pihl

Rasmussen

et al. 2019

Journal of Bone and Mineral Research

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Hypoparathyroidism (HP) is a condition of parathyroid hormone (PTH) deficiency leading to abnormal calcium and phosphate metabolism. The mainstay of therapy consists of vitamin D and calcium supplements, as well as adjunct Natpara (PTH(1‐84)). However, neither therapy optimally controls urinary calcium (uCa) or significantly reduces the incidence of hypercalcemia and hypocalcemia. TransCon PTH, a sustained‐release prodrug of PTH(1‐34) in development for the treatment of HP, was designed to overcome these limitations. To determine the pharmacokinetics and pharmacodynamics of TransCon PTH, single and repeat s.c. dose studies were performed in rats and monkeys. TransCon PTH demonstrated a half‐life of 28 and 34 hours in rats and monkeys, respectively. After repeated dosing, an infusion‐like profile of the released PTH, characterized by low peak‐to‐trough levels, was obtained in both species. In intact rats and monkeys, daily subcutaneous administration of TransCon PTH was associated with increases in serum calcium (sCa) levels and decreases in serum phosphate levels (sP). In monkeys, at a single dose of TransCon PTH that increased sCa levels within the normal range, a concurrent decrease in uCa excretion was observed. In 4‐week repeat‐dose studies in intact rats and monkeys, uCa excretion was comparable to controls across all dose levels despite increases in sCa levels. Further, in a rat model of HP, TransCon PTH normalized sCa and sP levels 24 hours per day. This was in contrast to only transient trends toward normalization of sCa and sP levels with an up to 6‐fold higher molar dose of PTH(1‐84). After repeated dosing to HP rats, uCa excretion transiently increased, corresponding to increases in sCa above normal range, but at the end of the treatment period, uCa excretion was generally comparable to sham controls. TransCon PTH was well tolerated and the observed pharmacokinetics and pharmacodynamics were in line with the expected action of physiological replacement of PTH. © 2019 American Society for Bone and Mineral Research.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Bioactivity Assaymentioning

confidence: 99%

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Design and Preclinical Development of TransCon PTH, an Investigational Sustained-Release PTH Replacement Therapy for Hypoparathyroidism

Holten-Andersen

Pihl

Rasmussen

et al. 2019

Journal of Bone and Mineral Research

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“…CV index obtained using 3 series of the sensibilized erythrocytes received in various experimental days from different animals. CV index should not exceed 25% for cell methods [12]. CV should not exceed 11.5% for different erythrocytes series (figure 4).…”

Section: правильность разработанного метода (Accuracy) оценивали по фmentioning

confidence: 97%

“…Устойчивость метода оценивали по ва-риабельности значений IC 50 для одной серии препарата, по показателю CV, ис-пользуя 3 серии сенсибилизированных эритроцитов, полученных в разные экспе-риментальные дни от разных животных. Для клеточных методов показатель CV не должен превышать 25.0% [12]. В экспери-ментах CV для разных партий эритроци-тов составило не более 11.5% (рис.…”

Section: правильность разработанного метода (Accuracy) оценивали по фunclassified

Method development and validation for determination of antihemolytic activity of eculizumab (Soliris)

Прудникова¹,

Порошин²,

Кудина³

et al. 2016

Farm. farmakol. (Pâtigorsk)

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Detection of parathyroid hormone using an electrochemical impedance biosensor based on PAMAM dendrimers

Özcan

Sezgintürk

2015

Biotechnology Progress

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This paper presents a novel hormone-based impedimetric biosensor to determine parathyroid hormone (PTH) level in serum for diagnosis and monitoring treatment of hyperparathyroidism, hypoparathyroidism and thyroid cancer. The interaction between PTH and the biosensor was investigated by an electrochemical method. The biosensor was based on the gold electrode modified by 12-mercapto dodecanoic (12MDDA). Antiparathyroid hormone (anti-PTH) was covalently immobilized on to poly amidoamine dendrimer (PAMAM) which was bound to a 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS) couple, self-assembled monolayer structure from one of the other NH2 sites. The immobilization of anti-PTH was monitored by electrochemical impedance spectroscopy, cyclic voltammetry and scanning electron microscope techniques. After the optimization studies of immobilization materials such as 12MDDA, EDC-NHS, PAMAM, and glutaraldehyde, the performance of the biosensor was investigated in terms of linearity, sensitivity, repeatability, and reproducibility. PTH was detected within a linear range of 10-60 fg/mL. Finally the described biosensor was used to monitor PTH levels in artificial serum samples.

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Development and validation of a novel cell-based assay for potency determination of human parathyroid hormone (PTH)

Cited by 9 publications

References 21 publications

Design and Preclinical Development of TransCon PTH, an Investigational Sustained-Release PTH Replacement Therapy for Hypoparathyroidism

Design and Preclinical Development of TransCon PTH, an Investigational Sustained-Release PTH Replacement Therapy for Hypoparathyroidism

Method development and validation for determination of antihemolytic activity of eculizumab (Soliris)

Detection of parathyroid hormone using an electrochemical impedance biosensor based on PAMAM dendrimers

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