高分辨率的离子移动性质谱仪用于分离寡核酸酸二二聚合物
Molly S Blevins1, Jie Du1,2, Ralph Aderorho1,3
1Synthetic Molecule Analytical Chemistry, Synthetic Molecule Pharmaceutical Sciences, Genentech Inc., South San Francisco, California 94080, United States.
Analytical chemistry
|January 28, 2026
概括
在寡核酸治疗中酸 (PS) 修改产生复杂的二聚体. 本研究系统地使用高分辨率离子流动性质谱法 (HRIM-MS) 分析PS二聚体分离,为治疗性寡核化物实现出色的分辨率.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 化学生物学 化学生物学
背景情况:
- 酸 (PS) 修饰在寡核酸治疗中至关重要.
- PS链接引入二聚体,使分析表征复杂化.
- 现有的离子流动性质谱法 (IM-MS) 对PS异构体的研究是有限的.
研究的目的:
- 通过使用无损离子操纵 (SLIM) 高分辨率离子移动性 (HRIM) 结构与高分辨率质谱学 (HRMS) 结合,全面分析酸 (PS) 异构体.
- 系统地研究影响PS二聚体分离的因素,包括电荷状态,引状态,寡核酸长度,序列组成和PS修饰.
- 建立一个详细的框架,以优化PS改性寡核酸的IM-MS分析.
主要方法:
- 使用SLIM-HRIM与HRMS相结合,用于模型聚-dT PS寡核酸的系统分析.
- 研究了各种参数 (电荷状态,引电状态,长度,序列,PS号/位置) 对分离的影响.
- 通过一系列的寡核酸长度 (5-35 nt) 实现了PS异体的高分辨率分离.
主要成果:
- 证明增加序列不对称性和较低的寡核酸m/z增强了移动性分离分辨率.
- 对于PS寡核酸,达到高达5.0的峰值至峰值分辨率 (Rpp),具有一致的基线分离.
- 确定了对 adduct 状态选择和潜在误解的关键见解.
- 成功地解决了治疗单导向RNA (sgRNA) 片段的双立体体.
结论:
- 开发了迄今为止最详细的框架,以优化PS改性寡核酸的IM-MS分析.
- 提供了关键的洞察力,对结构和方法因素,规范PS diastereomer分离提供了关键的见解.
- 强调了序列不对称性和 adduct 状态在实现最佳分离方面的重要性.
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