Sequencing of Phosphorodiamidate Morpholino Oligomers by Hydrophilic Interaction Chromatography Coupled to Tandem Mass Spectrometry.

IF 3.1 2区 化学 Q2 BIOCHEMICAL RESEARCH METHODS Journal of the American Society for Mass Spectrometry Pub Date : 2024-10-02 Epub Date: 2024-08-30 DOI:10.1021/jasms.4c00281
Mingming Wang, Brian O'Day, Brian Michaels, Jurjus Jurayj, Bao Zhong Cai, Tao Wei
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Abstract

Sequencing of phosphorodiamidate morpholino oligomers (PMOs) by hydrophilic interaction chromatography (HILIC) coupled to tandem mass spectrometry (MS/MS) is reported. The MS/MS analysis was performed using a quadrupole/time-of-flight (Q-ToF) mass analyzer and collision induced dissociation (CID) in negative ion mode. To improve MS sensitivity in negative ion mode, HILIC conditions, including the separation column, mobile phases, and MS parameters, were optimized. Using the developed HILIC-CID-MS/MS method, 100% sequence coverage was achieved for PMOs ranging from 18-mer to 25-mer. Additionally, the method was successfully applied to identifying positional isomers of n - 1 deletion impurities present in PMO drug substances.

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利用亲水相互作用色谱法和串联质谱法对磷酸二酰胺吗啉低聚物进行测序。
报告采用亲水相互作用色谱法(HILIC)结合串联质谱法(MS/MS)对磷酸二酰胺吗啉低聚物(PMOs)进行了测序。MS/MS 分析采用四极杆/飞行时间(Q-ToF)质量分析仪和负离子模式下的碰撞诱导解离(CID)。为了提高负离子模式下的质谱灵敏度,对包括分离柱、流动相和质谱参数在内的 HILIC 条件进行了优化。利用所开发的 HILIC-CID-MS/MS 方法,对 18-mer 至 25-mer 的 PMOs 实现了 100% 的序列覆盖。此外,该方法还成功应用于鉴定 PMO 药物中 n - 1 缺失杂质的位置异构体。
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来源期刊
CiteScore
5.50
自引率
9.40%
发文量
257
审稿时长
1 months
期刊介绍: The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role. Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives
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