Development of a robust reporter gene-based assay for the bioactivity determination of recombinant human follicle stimulating hormone (rhFSH) pharmaceutical products

• Published in: Journal of pharmaceutical and biomedical analysis 2020-01, Vol.177, p.112855-112855, Article 112855
• Main Authors: Wang, Lv-yin, Liang, Cheng-gang, Yang, Hui-min, Li, Jing, Li, Zhan-jun, Zhang, Hui, Lv, Ping, Li, Yi, Wang, Jun-zhi
• Format: Article
• Language: English
• Subjects: Animals; Bioactivity; Biological Assay - methods; Cell Line; CHO Cells; Cricetulus; Cyclic AMP - genetics; Feasibility Studies; Follicle stimulating hormone; Follicle Stimulating Hormone, Human - pharmacology; Genes, Reporter - genetics; In-vivo bioassay; Luciferases - chemistry; Luciferases - genetics; Ovarian weight gain; Receptors, FSH - genetics; Receptors, FSH - metabolism; Recombinant Proteins - pharmacology; Reporter gene assay; Response Elements - genetics; Validation
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2019.112855

[Display omitted] •A reporter gene assay (RGA) for bioactivity determination of FSH was established, optimized and validated.•The RGA can overcome the limitations of the in-vivo rat bioassay.•The RGA was not only highly consistent with the current in-vivo rat bioassay, but also superior on precision, accuracy and simplicity.•The RGA can be used as a powerful tool for the quality control and bio-similar drug comparability studies of FSH products. FSH plays a key role in the function of the reproductive system of human beings and is widely used both diagnostically and therapeutically in reproductive medicine. With the growing incidence of infertility, the demand for FSH pharmaceutical products is increasing. For this reason, the quality control process for FSH products is becoming more stringent. An accurate determination of bioactivity is crucial for the safety and efficacy of recombinant human follicle stimulating hormone (rhFSH). Up to now, in-vivo bioassay based on FSH-induced increases in rat ovarian weight has been the only method widely accepted by different pharmacopoeias. However this method has such drawbacks as the complex procedures, long assay period and high variability. Here, we established a reporter gene assay (RGA) based on the CHO-K1-FSHR-CRE-Luc cell line that stably expresses human follicle stimulating hormone receptor (hFSHR), as well as a luciferase reporter under the control of cyclic adenosine monophosphate (cAMP) response elements (CRES). Our study showed that our new assay not only has good dose-dependent responsiveness to rhFSH, but it also performs excellently in terms of specificity, precision, linearity, and simplicity compared with in-vivo rat bioassays. These results implied that this robust reporter gene assay may be a viable supplement to the animal in-vivo bioassay and may be employed in potency determination of rhFSH pharmaceutical products.