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Updated: Jun 18, 2026

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
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Purification of Double-Stranded RNA Impurities From In Vitro-Transcribed mRNA Using Size-Exclusion Chromatography.

Kyle J Tynan1,2, Trina Mouchahoir1,2, Mark S Lowenthal1

  • 1National Institute of Standards and Technology (NIST), Gaithersburg, Maryland, USA.

Biomedical Chromatography : BMC
|June 17, 2026
PubMed
Summary
This summary is machine-generated.

Double-stranded RNA (dsRNA) impurities are removed from messenger RNA (mRNA) using a novel size-exclusion chromatography method. This technique enhances vaccine efficacy by purifying mRNA therapeutics.

Keywords:
dsRNAhydrodynamic chromatographyin vitro transcriptionmRNApurificationsize‐exclusion chromatography

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Double-stranded RNA (dsRNA) is a byproduct of in vitro mRNA transcription.
  • dsRNA triggers immunogenic responses, reducing mRNA vaccine efficacy.
  • Separating dsRNA from mRNA is difficult due to similar physicochemical properties.

Purpose of the Study:

  • To develop an effective method for removing dsRNA impurities from mRNA.
  • To improve the purity and efficacy of mRNA therapeutics.

Main Methods:

  • A lab-scale size-exclusion chromatography (SEC) method was developed.
  • The SEC method was optimized for separating dsRNA from mRNA.

Main Results:

  • Baseline resolution between dsRNA and mRNA was achieved (resolution factor = 1.6).
  • High recovery of mRNA was maintained (81.8%).
  • The method effectively removes immunogenic dsRNA impurities.

Conclusions:

  • The developed SEC method provides a novel chromatographic strategy for purifying mRNA.
  • This purification strategy is crucial for enhancing the safety and efficacy of mRNA therapeutics.
  • This technique addresses a key challenge in the manufacturing of mRNA-based medicines.