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. 2025 Sep 18;27(6):144.
doi: 10.1208/s12248-025-01122-w.

Technical Considerations of Pharmacokinetic Assays for LNP-mRNA Drug Products by RT-qPCR

Jason Pennucci  1 , Amanda Hays  2 , Wendy Adamowicz  3 , Mitra Azadeh  4 , Mohamed Benhammadi  5 , Enric Bertran Portabella  6 , MingLai Cheng  7 , Kelly Colletti  8 , Sanjay L Dholakiya  9 , Rajitha Doddareddy  10 , Lara Duchstein  11 , Jennifer Durham  12 , Sebastian Guelman  13 , Bryan Gullick  14 , Kate Herr  15 , Darshana Jani  1 , Niketa Jani  16 , Oskar Johansson  17 , Kaarthik John  18 , Mikael Kubista  19 , Amy Lavelle  3 , Geng Li  20 , Yanchun Li  15 , Hsing-Yin Liu  15 , Timothy Lochmann  3 , Haiyan Ma  21 , Benjamin Moritz  22 , Jacqueline Murphy  15 , Alice Park  9 , Suresh Peddigari  1 , Marie-Soleil Piche  5 , Swarna Suba Ramaswamy  23 , Asha Reddy  1 , Muriel Smet  24 , Johannes Stanta  25 , Jenny Valentine  26 , Katherine Veirs  7 , Venkata Vepachedu  27 , Mark Wissel  28 , Meizhen Wu  15 , Bin Xu  29 , Susan Zondlo  7
Affiliations

Technical Considerations of Pharmacokinetic Assays for LNP-mRNA Drug Products by RT-qPCR

Jason Pennucci et al. AAPS J. .

Abstract

Lipid nanoparticle-messenger RNA (LNP-mRNA) drug products are a growing class of drug modalities. The unique composition of these drug products requires multiple measurements to account for the different components of these drug modalities. Pharmacokinetic (PK) measurements include measurement of the encapsulated mRNA and components of the LNP in circulation to understand the effectiveness of the therapeutic mRNA. The PK measurements can utilize many different platforms including PCR. Current regulatory guidance documents for bioanalytical method validation are specific to ligand binding and chromatographic assay methods and difficult to interpret for use with molecular workflows. The purpose of this paper is to provide information on considerations for validation of regulated reverse transcription quantitative PCR (RT-qPCR) assays that are used to support the pharmacokinetic analysis of LNP-mRNA drug products.

Keywords: LNP; MRNA; PK; RT-dPCR; RT-qPCR.

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Conflict of interest statement

Declarations. Conflict of interest: The authors are employed by and receive compensation from their affiliations that are listed in this manuscript. The authors have no other relevant affiliations or financial involvements with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript. Disclaimer: The views and conclusions presented in this paper are those of the authors and do not necessarily reflect the representative affiliation or company’s position on the subject.

References

    1. Sparmann A, Vogel J. RNA-based medicine: from molecular mechanisms to therapy. EMBO J. 2023;42(21):e114760. https://doi.org/10.15252/embj.2023114760 . - DOI - PubMed - PMC
    1. Xu S, Yang K, Li R, Zhang L. mRNA vaccine era-mechanisms, drug platform and clinical prospection. Int J Mol Sci. 2020;21(18):6582. https://doi.org/10.3390/ijms21186582 . - DOI - PubMed - PMC
    1. Kon E, Ad-El N, Hazan-Halevy I, Stotsky-Oterin L, Peer D. Targeting cancer with mRNA-lipid nanoparticles: key considerations and future prospects. Nat Rev Clin Oncol. 2023;20(11):739–54. https://doi.org/10.1038/s41571-023-00811-9 . - DOI - PubMed
    1. Sahin U, Karikó K, Türeci Ö. mRNA-based therapeutics—developing a new class of drugs. Nat Rev Drug Discov. 2014;13(10):759–80. - DOI - PubMed
    1. Szabo GT, Mahiny AJ, Vlatkovic I. COVID-19 mRNA vaccines: platforms and current developments. Mol Ther. 2022;30(5):1850–68. https://doi.org/10.1016/j.ymthe.2022.02.016 . - DOI - PubMed - PMC

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