ArticleScientific reports2019
HPLC methods for purity evaluation of man-made single-stranded RNAs.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
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.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
30 citing papers in PubMed, 92 citations in OpenAlex.
- Advances in Analysis of Therapeutic Oligonucleotides with Chromatography Coupled to Mass Spectrometry.Analytical chemistry · 2026Review
- Synthesis of long and functionally active RNAs facilitated by acetal levulinic ester chemistry.Nucleic acids research · 2026Article
- Challenges and advances in analytical separation techniques for RNA-lipid nanoparticle therapeutics.Analytical and bioanalytical chemistry · 2026Review
- Coumarin-Based Photolabile Solid Support Facilitates Nonchromatographic Purification of RNA Oligonucleotides.The Journal of organic chemistry · 2025Article
- The Potential and Limitations of the MinION/Yenos Platform for miRNA-Enabled Early Cancer Detection.International journal of molecular sciences · 2025Article
- Current Analytical Strategies for mRNA-Based Therapeutics.Molecules (Basel, Switzerland) · 2025Review
- Characterisation and analysis of mRNA critical quality attributes using liquid chromatography based methods.Journal of chromatography. A · 2025Review
- High-Throughput Quantification and Characterization of Dual Payload mRNA/LNP Cargo via Deformulating Size Exclusion and Ion Pairing Reversed Phase Assays.Analytical chemistry · 2025Article
- High Sensitivity and Specificity Platform to Validate MicroRNA Biomarkers in Cancer and Human Diseases.Non-coding RNA · 2024Article
- Unravelling the Link between Oligonucleotide Structure and Diastereomer Separation in Hydrophilic Interaction Chromatography.Analytical chemistry · 2024Article
- Nanotherapeutic Heterogeneity: Sources, Effects, and Solutions.Small (Weinheim an der Bergstrasse, Germany) · 2024Review
- Digital Twin Fundamentals of mRNAACS omega · 2024Article
- Anion exchange HPLC monitoring of mRNAFrontiers in molecular biosciences · 2024Article
- Liquid Chromatography Methods for Analysis of mRNA Poly(A) Tail Length and Heterogeneity.Analytical chemistry · 2023Article
- High Recovery Chromatographic Purification of mRNA at Room Temperature and Neutral pH.International journal of molecular sciences · 2023Article
- Light-Up Split Broccoli Aptamer as a Versatile Tool for RNA Assembly Monitoring in Cell-Free TX-TL Systems, Hybrid RNA/DNA Origami Tagging and DNA Biosensing.International journal of molecular sciences · 2023Article
- Challenges and emerging trends in liquid chromatography-based analyses of mRNA pharmaceuticals.Journal of pharmaceutical and biomedical analysis · 2023Review
- The Storage and In-Use Stability of mRNA Vaccines and Therapeutics: Not A Cold Case.Journal of pharmaceutical sciences · 2023Review
- High-throughput iSpinach fluorescent aptamer-based real-time monitoring of in vitro transcription.Bioresources and bioprocessing · 2022Article
- A High-Pressure, High-Temperature Flow Reactor Simulating the Hadean Earth Environment, with Application to the Pressure Dependence of the Cleavage of Avocado Viroid Hammerhead Ribozyme.Life (Basel, Switzerland) · 2022Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
1 author at 1 institution in 1 country.
Funding
Abstract
Synthetic RNA oligos exhibit purity decreasing as a function of length, because the efficiency of the total synthesis is the numerical product of the individual step efficiencies, typically below 98%. Analytical methods for RNAs up to the 60 nucleotides (nt) have been reported, but they fall short for purity evaluation of 100nt long, used as single guide RNA (sgRNA) in CRISPR technology, and promoted as pharmaceuticals. In an attempt to exploit a single HPLC method and obtain both identity as well as purity, ion-pair reversed-phase chromatography (IP-RP) at high temperature in the presence of an organic cosolvent is the current analytical strategy. Here we report that IP-RP is less suitable compared to the conventional ion-exchange (IEX) for analysis of 100nt RNAs. We demonstrate the relative stability of RNA in the denaturing/basic IEX mobile phase, lay out a protocol to determine the on-the-column stability of any RNA, and establish the applicability of this method for quality testing of sgRNA, tRNA, and mRNA. Unless well resolving HPLC methods are used for batch-to-batch evaluation of man-made RNAs, process development will remain shortsighted, and observed off-target effects in-vitro or in-vivo may be partially related to low purity and the presence of shorter sequences.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.