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解码复杂性在合成的寡核酸:解凝聚异酸不纯离子通过高分辨率质谱学解
A M Abdullah1, Cynthia Sommers1, Jason D Rodriguez1
1Division of Complex Drug Analysis, Office of Testing and Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, St. Louis, Missouri 63110, United States.
Analytical chemistry
|December 29, 2023
概括
本研究引入了一种使用高分辨率质谱的直接方法,用于识别和量化合成寡核酸中的凝结杂质. 该技术利用同位素包裹进行核酸变异的准确分析,改进了杂质分析.
科学领域:
- 分析化学 分析化学
- 质谱测量质量谱测量
- 氧核酸治疗药物 治疗药物
背景情况:
- 液体染色学-质谱学 (LC-MS) 由于质量相似,难以将合成寡核酸中的凝聚杂质分离出来.
- 诸如 uracil 或cytosine 核酸的添加/删除之类的杂质会给分析带来重大挑战.
- 现有的方法没有能力量化异杂质离子内的多种序列变异.
研究的目的:
- 开发一种直接方法来识别和量化合成寡核酸中异酸杂质离子.
- 为了克服传统LC-MS用于分析复杂杂质混合物的局限性.
- 为了能够在寡核酸治疗中准确地进行杂质分析.
主要方法:
- 使用高分辨率质谱法 (HRMS) 来分辨杂质离子的同位素包裹.
- 使用完整或有针对性的MS分析来测量化学配方依赖的同位素分布.
- 使用测量或理论调整的同位素分布,绕过标准曲线,量化同位素杂质离子.
主要成果:
- 通过核酸添加或删除成功识别和定量化等离子杂质离子.
- 证明了在单个杂质离子类型中量化多个序列变异的能力.
- 验证了理论同位素分布的使用,在没有标准的情况下使用校正因数.
结论:
- 基于HRMS的方法有效地解决了合成寡核酸中凝结的异杂质.
- 这种方法为深入的杂质分析提供了可靠的解决方案,这对于核酸治疗药物开发至关重要.
- 与传统的MS/MS方法相比,该技术为复杂杂质混合物提供了增强的量化能力.
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