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CC BY 4.0 · Pharmaceutical Fronts 2024; 06(03): e315-e321
DOI: 10.1055/s-0044-1788781
Original Article

Development, Optimization, and Validation of an in vitro Cell-Based Bioassay to Determine the Biological Activity of Teriparatide (PTH1–34)

Tao Luo
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
,
Jianguang Lu
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
2   Shanghai Duomirui Biotechnology Co., Ltd., Shanghai, People's Republic of China
,
Chen Guo
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
3   School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People's Republic of China
,
Xue Feng
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
,
Jun Xu
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
2   Shanghai Duomirui Biotechnology Co., Ltd., Shanghai, People's Republic of China
,
Jun Feng
1   China State Institute of Pharmaceutical Industry, Shanghai, People's Republic of China
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Abstract

This study aimed to establish an efficient in vitro cell-based assay to measure the activity of teriparatide (PTH1–34). In this study, a rat osteosarcoma cell line (UMR-106) was treated with various concentrations of PTH1–34, and the biological activity of PTH1–34 was determined by quantitatively measuring intracellular cyclic adenosine monophosphate levels using a time-resolved fluoroimmunoassay. A four-parameter fitting analysis was used to calculate the relative potency of the samples. The experimental conditions were optimized. The method's specificity, relative accuracy, precision, and linearity were validated. Our data suggested that this method had good specificity, a relative bias of relative accuracy ranging from −0.8 to 1.4%, a correlation coefficient for the linear regression equation of 0.9953, a geometric coefficient of variation for intermediate precision ranges from 2.0 to 3.5%, and a linear range of 50 to 150%. This method significantly improves the quality control and release inspection efficiency of PTH1–34 and may be further developed and validated as an alternative to the existing United States Pharmacopeia and European Pharmacopoeia inclusion methods. This method also provides a platform for the high-throughput screening of PTH1–34 analogs.

Keywords

teriparatide - in vitro bioassay - cAMP - time-resolved fluoroimmunoassay - method verification


Publikationsverlauf

Eingereicht: 09. Januar 2024

Angenommen: 15. Juli 2024

Artikel online veröffentlicht:
06. August 2024

© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

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