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

被引：0
|
作者：
Wang, Mingming [1 ]
O'Day, Brian [1 ]
Michaels, Brian [1 ]
Jurayj, Jurjus [1 ]
Cai, Bao Zhong [1 ]
Wei, Tao [1 ]
机构：
[1] Sarepta Therapeut Inc, Cambridge, MA 02142 USA
来源：
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY | 2024年 / 35卷 / 10期
关键词：
tandem mass spectrometry (MS/MS); collision induceddissociation (CID); hydrophilic interaction chromatography(HILIC); phosphorodiamidate morpholino oligomer (PMO); sequencing; impurity characterization; PHOSPHOROTHIOATE OLIGONUCLEOTIDES; LIQUID-CHROMATOGRAPHY; ANTISENSE OLIGOMERS; INHIBITION; VIRUS; RNA; FRAGMENTATION; RESISTANCE; DNA;
D O I：
10.1021/jasms.4c00281
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：2490 / 2498
页数：9
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