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Assay for rapid quantification of capped and tailed intact mRNA

Rachel Y Gao1, Tianjing Hu1, Amber W Taylor1

  • 1InDevR Inc., 6035 Longbow Drive, Suite 102, Boulder, CO 80301, United States of America.

Vaccine
|June 4, 2025
PubMed

Insights

A new 5' CapQ assay offers rapid, accurate measurement of intact messenger RNA (mRNA), simplifying quality control for mRNA vaccine and therapeutic development. This assay streamlines analysis, reducing bottlenecks in critical bioprocessing steps.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Vaccine Development

Background:

  • Messenger RNA (mRNA) technology has advanced significantly, particularly for vaccines and therapeutics.
  • Assessing mRNA quality (integrity, 5' capping, 3' polyadenylation) is crucial but challenging with existing methods.
  • Current analytical techniques often involve lengthy procedures, complex preparation, and require specialized expertise.

Purpose of the Study:

  • To introduce a novel, rapid, and straightforward assay for evaluating mRNA quality.
  • To provide a single, unique measurement for intact mRNA, assessing both 5' cap and 3' poly(A) tail.
  • To overcome the limitations of current mRNA analytical methods.

Main Methods:

  • Development of the 5' CapQ assay, a benchtop-based method.
  • The assay performs quantitative or relative analysis of mRNA quality.
  • Analysis is completed within a 2-hour timeframe with minimal sample processing.

Main Results:

  • The 5' CapQ assay provides accurate and precise measurements of intact mRNA.
  • It simplifies the assessment of mRNA integrity, 5' capping, and 3' polyadenylation.
  • The assay demonstrates high performance without extensive sample preparation.

Conclusions:

  • The 5' CapQ assay represents a significant advancement in mRNA analytics.
  • It effectively alleviates bottlenecks in optimizing in vitro transcription (IVT) and capping processes.
  • The assay facilitates consistent assessment of batch-to-batch quality in mRNA bioprocessing.
Keywords:
5′ capping efficiencyIntegrityVaccinemRNA

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