ArticleAnalytical chemistry2026
Automated Online Direct mRNA Sequence Mapping Using Partial RNase T1 Digests.
Article in Analytical chemistry, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
2 citing papers in PubMed.
- Human 28S rRNA analysed by state-of-the-art oligonucleotide mass spectrometry: benchmarking current capabilities and a call to action for MS-Seq.bioRxiv : the preprint server for biology · 2026Article
- A nanogram-sensitive workflow for oligonucleotide mass spectrometry using ion-pair-free nanoflow HILIC and RNase benchmarking.Nucleic acids research · 2026Article
Corrections and comments
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Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mass spectrometry-based approaches have emerged as powerful tools for the analysis of a wide range of critical quality attributes of mRNA medicines, including sequence identity, 5' capping efficiency, and 3' poly(A) tail length and heterogeneity. These critical quality attributes can impact the quality, safety, and efficacy of mRNA medicines. In this study, we have utilized online partial RNase T1 digests in conjunction with two-dimensional liquid chromatography mass spectrometry (2D LC-MS) for the direct sequence mapping of mRNA. Automated online partial RNase T1 digests are performed in conjunction with ion-pair reversed-phase liquid chromatography. No sample or solvent manipulation is required following online RNase digestions, demonstrating the simplicity of the method. High-resolution tandem mass spectrometry was used to identify the corresponding oligoribonucleotides and generate mRNA sequence maps. High sequence coverage (93-99%) for eGFP mRNA was obtained in <60 min based only on unique oligoribonucleotide identifications. Moreover, the online partial RNase T1 digests result in controlled, fully automated and reproducible mRNA digests, enabling high-throughput, direct mRNA sequence mapping studies. The online partial RNase digests offer significant advantages over existing methods for rapid, automated mRNA identity testing. Furthermore, precise control of the digest conditions via flow rate and temperature of the online RNase T1 digest, enables multiattribute monitoring of 5' capping efficiency, mRNA sequence mapping, and 3' poly(A) tail length and heterogeneity in a fully automated 2D LC-MS workflow.
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Registered trials
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