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Quantification of mRNA in Lipid Nanoparticles Using Mass Spectrometry.

Mark S Lowenthal1, Abigail S Antonishek1, Karen W Phinney1

  • 1Biomolecular Measurement Division, Material Measurement Lab, National Institute of Standards and Technology (NIST), Gaithersburg, Maryland 20899, United States.

Analytical Chemistry
|January 8, 2024
PubMed
Summary

Accurate quantification of lipid nanoparticle-encapsulated mRNA (LNP-mRNA) therapeutics is crucial for safety. A novel acid hydrolysis liquid chromatography-mass spectrometry (LC-MS) method provides traceable mRNA quantification without extraction, ensuring reliable therapeutic product analysis.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Pharmaceutical Science

Background:

  • Lipid nanoparticle-encapsulated mRNA (LNP-mRNA) represents a promising therapeutic modality for various diseases.
  • Accurate quantification of mRNA in LNP-mRNA products is essential for ensuring therapeutic consistency and patient safety.
  • Measurement traceability is a critical factor in achieving accurate quantity determination.

Purpose of the Study:

  • To develop and validate a novel method for traceable quantification of mRNA within LNP-mRNA formulations.
  • To establish a sample preparation technique that avoids complex extraction procedures, detergents, or enzymes.
  • To demonstrate the applicability of the method for quantifying mRNA in both active substances and formulated drug products.

Main Methods:

  • Development of a liquid chromatography-mass spectrometry (LC-MS) approach for in situ LNP-mRNA measurement.
  • Utilizing direct acid hydrolysis of LNP-mRNA products followed by an isotope dilution strategy.
  • Establishing traceability through common nucleobase calibrators for oligonucleotide quantification.

Main Results:

  • The developed LC-MS method enables accurate and traceable quantification of mRNA in LNP-mRNA products.
  • The acid hydrolysis sample preparation is efficient, bypassing the need for mRNA extraction, detergents, or enzymes.
  • The method provides quantitative analysis of mRNA that is independent of time and location, ensuring robustness.

Conclusions:

  • Acid hydrolysis coupled with LC-MS offers a reliable and traceable method for quantifying mRNA in LNP-mRNA therapeutics.
  • This approach simplifies sample preparation, enhancing the efficiency and accuracy of quality control for mRNA drug products.
  • The validated method supports the development and consistent manufacturing of safe and effective LNP-mRNA-based therapies.