Application of ion mobility-mass spectrometry to microRNA analysis(METHODS)
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概要
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Liquid chromatography/mass spectrometry is widely used for studying sequence determination and modification analysis of small RNAs. However, the efficiency of liquid chromatography-based separation of intact small RNA species is insufficient, since the physiochemical properties among small RNAs are very similar. In this study, we focused on ion mobility-mass spectrometry (IM-MS), which is a gas-phase separation technique coupled with mass spectrometry; we have evaluated the utility of IM-MS for microRNA (miRNA) analysis. A multiply charged deprotonated ion derived from an 18-24-nt-long miRNA was formed by electrospray ionization, and then the time, called the "drift time", taken by each ion to migrate through a buffer gas was measured. Each multivalent ion was temporally separated on the basis of the charge state and structural formation; 3 types of unique mass-mobility correlation patterns (i.e., chainlike-form, hairpin-form, and dimer-form) were present on the two-dimensional mobility-mass spectrum. Moreover, we found that the ion size (sequence length) and the secondary structures of the small RNAs strongly contributed to the IM-MS-based separation, although solvent conditions such as pH had no effect. Therefore, sequence isomers could also be discerned by the selection of each specific charged ion, i.e., the 6^- charged ion reflected a majority among chainlike-, hairpin-, and other structures. We concluded that the IM-MS provides additional capability for separation; thus, this analytical method will be a powerful tool for comprehensive small RNA analysis.
- 公益社団法人日本生物工学会の論文
- 2013-03-00
著者
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Bamba Takeshi
Department Of Applied Environmental Biology Graduate School Of Pharmaceutical Sciences Osaka Univers
-
Fukusaki Eiichiro
Department Of Biotechnology Faculty Of Engineering Osaka University
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Izumi Yoshihiro
Department Of Biological Science Faculty Of Science Kumamoto University
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Hirose Kenji
Nihon Waters K.K.
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Takebayashi Kosuke
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Izumi Yoshihiro
Department of Internal Medicine, Graduate School of Medicine, Kobe University
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