School of Life Science and Biopharmaceuticals, Shenyang Pharmaceutical University, No. 103 Wenhua Road, Shenyang, Liaoning 110016, China; Shenyang Sunshine Pharmaceutical CO., Ltd, 1(st) 3, 10(th) Road, Economic and Technological Development Zone, Shenyang, Liaoning 110027, China.
Shenyang Sunshine Pharmaceutical CO., Ltd, 1(st) 3, 10(th) Road, Economic and Technological Development Zone, Shenyang, Liaoning 110027, China.
Int Immunopharmacol. 2021 Oct;99:107982. doi: 10.1016/j.intimp.2021.107982. Epub 2021 Jul 29.
Recombinant human thrombopoietin (rhTPO) was approved by the National Medical Products Administration in 2010 for the treatment of thrombocytopenia in patients with immune thrombocytopenic purpura and chemotherapy-induced thrombocytopenia. Nevertheless, no method for determining rhTPO bioactivity has been recorded in different national/regional pharmacopoeia. Novel methods for lot release and stability testing are needed that are simpler, quicker, and more accurate. Here, we developed a novel reporter gene assay (RGA) for rhTPO bioassay with Ba/F3 cell lines that stably expressed human TPO receptor and luciferase reporter driven by sis-inducible element, gamma response region, and gamma-interferon activated sequence. During careful optimization, the RGA method demonstrated high performance characteristics. According to the International Council for Harmonization Q2 (R1) guidelines and the Chinese Pharmacopoeia 2020 edition, the validation results demonstrated that this method is highly time-saving, sensitive, and robust for research, development, manufacture, and quality control of rhTPO.
2010 年，国家药品监督管理局批准重组人血小板生成素（rhTPO）用于治疗免疫性血小板减少症和化疗引起的血小板减少症患者的血小板减少症。然而，不同国家/地区的药典中都没有记录 rhTPO 生物活性的测定方法。需要开发更简单、更快速、更准确的新型放行和稳定性测试方法。在这里，我们开发了一种新型的报告基因检测（RGA），用于 Ba/F3 细胞系中 rhTPO 的生物测定，该细胞系稳定表达人 TPO 受体和由 sis 诱导元件、γ反应区和γ干扰素激活序列驱动的荧光素酶报告基因。在仔细优化过程中，RGA 方法表现出了高性能特征。根据国际协调会议 Q2（R1）指南和 2020 年版中国药典，验证结果表明，该方法对于 rhTPO 的研究、开发、制造和质量控制具有高度省时、灵敏和稳健的特点。
