在复杂的生物样本中自动识别寡核酸代谢物,使用超高性能液体染色学高分辨率质谱学与分子分析软件相结合
Xinyuan Ye1, Lili Jiang1, Ying Xiong1
1Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030, China.
概括
本研究引入了一种自动化策略,用于识别使用UHPLC-QTOF-MS/MS和软件的寡核酸 (ON) 代谢物. 该方法成功地在体外和体内鉴定了17种Fomivirsen代谢物,主要是通过3'-外核酶.
科学领域:
- 生物治疗学 生物治疗学
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
背景情况:
- 氧核酸 (ON) 疗法是一个快速发展的领域.
- 超高性能液体染色学高分辨率质谱 (UHPLC-HRMS) 对于代谢物识别至关重要.
- 用于ON代谢物识别的手动数据处理是耗时的.
研究的目的:
- 建立和验证用于识别寡核酸代谢物的自动化分析策略.
- 简化和加快ON代谢物识别过程.
- 调查福米维尔森 (FMVS) 的体外和体内代谢.
主要方法:
- 使用UHPLC-QTOF-MS/MS和分子分析软件进行自动代谢物识别的分析策略的开发.
- 优化样品准备和UHPLC-QTOF-MS/MS条件的优化.
- 使用Fomivirsen (FMVS) 的酶消化剂验证该策略.
主要成果:
- 自动化策略成功识别了FMVS.的酶性消化物.
- 在血清,血和组织中共发现了17种FMVS代谢物.
- 福米维尔森主要由血,肝脏和脏中的3-外核酶代谢.
结论:
- 建立的策略可以自动识别寡核酸代谢产物.
- 这种方法大大简化了ON代谢物的识别过程.
- 该策略适用于识别其他寡核酸代谢产物.
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