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Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA
Published on: February 1, 2019
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Characterization of mRNA therapeutics
Guilherme J Guimaraes1, Jaeah Kim1, Michael G Bartlett1
1Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia, USA.
Mass Spectrometry Reviews
|July 4, 2023
Summary
Analytical methods are crucial for characterizing therapeutic messenger RNAs (mRNAs), enabling robust quality, identity, and integrity assessments for clinical applications. This review details techniques for mRNA analysis, supporting development and deployment.
Area of Science:
- Biotechnology and Pharmaceutical Sciences
- Molecular Biology and Genetics
Background:
- Therapeutic messenger RNAs (mRNAs) represent a significant advancement in treating complex diseases, owing to their protein-encoding capabilities.
- The large molecular size of therapeutic mRNAs, while beneficial for function, presents substantial analytical challenges.
- Standardized and advanced analytical methods are essential for the development and clinical progression of mRNA-based therapeutics.
Purpose of the Study:
- To review and discuss current analytical methodologies for characterizing therapeutic messenger RNA (mRNA).
- To evaluate the advantages and limitations of various techniques used for assessing mRNA quality, identity, and integrity.
- To highlight the application of these methods in critical quality attribute measurements, including capping efficiency, poly(A) tail length, and stability.
Main Methods:
- Gel electrophoresis for RNA size and integrity assessment.
- Liquid chromatography-mass spectrometry (LC-MS) for detailed molecular profiling.
- Next-generation sequencing (NGS) approaches, including shotgun sequencing, for comprehensive analysis.
- Intact mass measurements for precise molecular weight determination.
Main Results:
- A comprehensive overview of analytical techniques applicable to therapeutic mRNA characterization is presented.
- The review details the utility of methods for evaluating capping efficiency and poly(A) tail length, crucial for mRNA function and stability.
- Applicability of these analytical methods in assessing mRNA stability under various conditions is discussed.
Conclusions:
- Robust analytical characterization is indispensable for ensuring the quality, safety, and efficacy of therapeutic mRNAs.
- The discussed methods provide a foundation for the rigorous evaluation of therapeutic mRNA candidates.
- Continued development and validation of analytical techniques will further support the clinical translation of mRNA therapeutics.
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