Sannes-Lowery Kristin A, Hofstadler Steven A
Ibis Therapeutics, A Division of Isis Pharmaceuticals, Carlsbad, California 92008, USA.
J Am Soc Mass Spectrom. 2003 Aug;14(8):825-33. doi: 10.1016/S1044-0305(03)00335-0.
Modified oligonucleotides continue to play an important role as antisense compounds that inhibit the expression of genes associated with metabolic disorders, cancer, and infectious diseases. Because the majority of modifications render these molecules refractory to standard enzymatic sequencing techniques, alternative sequencing methods which are fast and reliable are needed. In this work we explore how sugar and backbone modifications affect fragmentation patterns observed from oligonucleotides which are fragmented by infrared multiple photon dissociation in the external reservoir of an electrospray ionization Fourier transform ion cyclotron mass spectrometer. The modifications influence which fragment types (i.e., a(n)-B versus c(n)) dominate and the ease with which the oligonucleotides are fragmented. General observations for confirming the sequence of oligonucleotides are described.
修饰寡核苷酸作为反义化合物，在抑制与代谢紊乱、癌症和传染病相关基因的表达方面继续发挥着重要作用。由于大多数修饰使这些分子难以用标准酶促测序技术进行测序，因此需要快速可靠的替代测序方法。在这项工作中，我们探讨了糖基和主链修饰如何影响寡核苷酸的片段化模式，这些寡核苷酸是在电喷雾电离傅里叶变换离子回旋共振质谱仪的外部储存器中通过红外多光子解离进行片段化的。这些修饰影响哪种片段类型（即a(n)-B与c(n)）占主导地位以及寡核苷酸片段化的难易程度。描述了用于确认寡核苷酸序列的一般观察结果。
