使用电子捕获解离质谱法对Morpholino反意义寡核酸进行测序
Kaoru Karasawa1, Eva Duchoslav2, Lyle Burton2
1SCIEX, 4-7-35 Kitashinagawa, Shinagawa, Tokyo 140-0001, Japan.
Analytical chemistry
|October 23, 2023
概括
我们使用电子捕获解离质谱测量对二胺酸盐形态寡合物 (PMOs) 进行了测序. 这种方法实现了100%的序列覆盖,使PMO杂质的有效分析成为可能.
科学领域:
- 分析化学 分析化学
- 质谱测量质量谱测量
- 橄核酸的化学成分
背景情况:
- 二胺酸形态寡合物 (PMO) 在反感性技术中至关重要.
- 精确的PMO测序对于质量控制和杂质分析至关重要.
- 现有的测序方法,如碰撞诱导解离,在提供完整的序列信息方面存在局限性.
研究的目的:
- 报告使用电子捕获解离 (ECD) 质谱法首次成功对PMO进行测序.
- 评估ECD在PMO分析中的效率和适用性.
- 开发一种工作流程,用于识别PMO杂质中的修饰点.
主要方法:
- 电子捕获解离 (ECD) 质谱法用于PMO测序.
- 分析的重点是18-25个mer的PMO.
- 建立了一个液体色谱-电子捕获解离并联质谱 (LC-ECD-MS/MS) 工作流.
主要成果:
- ECD以主要片段的形式生成d和z离子,从而获得100%的序列覆盖率.
- 这与碰撞诱导的解离形成鲜明对比,这种解离主要导致基损失.
- 一个通用的电子束能量 (15 eV) 在各种PMO序列,长度和电荷状态中有效.
- ECD反应速度与高性能液态染色学 (HPLC) 兼容.
结论:
- ECD质谱是一种高效的方法来测序PMOs,具有完整的序列覆盖.
- 通用能量设置简化了LC-MS/MS中的多前体准.
- 开发的LC-ECD-MS/MS工作流适用于测量PMO杂质中的修饰点.
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