A High-Throughput Workflow for Mass Spectrometry Analysis of Nucleic Acids by Nanoflow Desalting.
Michael B LanzillottiJennifer S BrodbeltPublished in: Analytical chemistry (2024)
Broad interest in nucleic acids, both their therapeutic capabilities and understanding the nuances of their structure and resulting function, has increased in recent years. Post-transcriptional modifications, in particular, have become an important analysis target, as these covalent modifications to the sugars, nitrogenous bases, and phosphate backbone impart differential functionality to synthetic and biological nucleic acids. Characterizing these post-transcriptional modifications can be difficult with traditional sequencing workflows; however, advancements in top-down mass spectrometry address these challenges. Online desalting platforms have enabled facile sample cleanup and reliable ionization of increasingly large (100 nt) oligonucleotides, and application of existing tandem mass spectrometry techniques has yielded information-rich spectra which can be used to interrogate primary sequences. To extend the capabilities of top-down MS and its analysis of nucleic acids, we have developed a nanoflow desalting platform for high-throughput and low sample-use desalting coupled with collision-induced dissociation (CID), 213 nm ultraviolet photodissociation (UVPD), and activated-ion electron photodetachment dissociation (a-EPD) to yield high-quality MS/MS spectra. Fragments identified using an m / z -domain isotope matching strategy yielded high sequence coverage (>70%) of a yeast phenylalanine tRNA.
Keyphrases
- gas chromatography
- high throughput
- mass spectrometry
- tandem mass spectrometry
- liquid chromatography
- liquid chromatography tandem mass spectrometry
- ultra high performance liquid chromatography
- solid phase extraction
- high performance liquid chromatography
- simultaneous determination
- ms ms
- single cell
- high resolution mass spectrometry
- high resolution
- electron transfer
- gene expression
- capillary electrophoresis
- health information
- transcription factor
- high glucose
- density functional theory
- social media
- electronic health record
- highly efficient
- quantum dots
- endothelial cells
- gold nanoparticles
- molecular dynamics
- affordable care act
- cell wall
- data analysis