Rapid and High-Confidence Sequence Confirmation of Modified Oligonucleotides using High Resolution Mass Spectrometry

摘要

Comparing to conventional methods such as chemical digestion, mass spectrometry has shown great potential for characterization of oligonucleotides, especially those with complex modifications. However, it is still challenging to achieve 100% sequence coverage using MS due to the increased complexity and difficulties for fragmentation of oligonucleotides with increased number of bases in the sequence. In this work, we developed a new method based on high resolution mass spectrometry for thorough fragmentation of highly modified oligonucleotides (～20 nucleotides in length), allowing rapid sequence confirmation with high confidence. Our method can determine the accurate mass of oligonucleotides and acquire abundant fragments within minutes, ensuring 100% sequence coverage of the target modified oligonucleotide.