开发和描述一种方法来确定人体复合叶状氨酸β子单元的N端异质性
Xinyue Hu1, Xuezhi Han2, Yue Sun1
1National Institutes for Food and Drug Control, Beijing, China.
Journal of pharmaceutical and biomedical analysis
|March 26, 2025
概括
一种新方法准确地测量了复合人卵泡刺激激素 (rhFSH) β子单元中的N端异质性. 这种基映射方法可确保生物类似rhFSH产品的质量控制.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 药理学 药理学是指药理学的学科.
背景情况:
- 重组人卵泡刺激激素 (rhFSH) 在生育治疗中至关重要.
- 在rhFSHβ子单元中的N端异质性可能会影响产品的有效性和安全性.
- 评估这种异质性的现有方法是有限的.
研究的目的:
- 开发和验证一种全面的方法来评估rhFSHβ子单元的N端异质性.
- 提高rhFSH生物类似产品的质量控制和可比性.
主要方法:
- 开发一种优化的片映射方法.
- 样品制备的整合,素消化和PNGase F脱糖化.
- 使用逆相高性能液态染色学 (RP-HPLC) 进行分离.
- 使用双重质谱法 (MS/MS) 确认了峰值的同一性.
主要成果:
- 成功分离集成 (NSCELTNITIAIEK) 和截断 (CELTNITIAIEK) 的βL1形式.
- 优化样品准备,将处理时间缩短到一天.
- 在多次评估中证明了稳定性和可重复性.
- 始终观察到来自不同制造商的β子单元中~50%的N端截断.
结论:
- 开发的方法为评估rhFSHβ子单元N终端异质性提供了强大的和高效的工作流.
- 这有助于提高质量控制,确保rhFSH生物仿制品的批量一致性和可比性.
- 改进的分析方法有助于rhFSH治疗的安全性和有效性.
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