ArticleThe AAPS journal2025
Comparison of Branched DNA to RT-qPCR Technology for the Quantitation of mRNA from mRNA-LNP Drug Product in Human Serum.
Article in The AAPS journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Accurate quantitation of circulating messenger RNA (mRNA) is critical for the quantitation of lipid nanoparticle-formulated mRNA (mRNA-LNP) drug products. This study evaluated the concordance between branched DNA (bDNA) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays for quantifying mRNA in human serum from a phase 1 clinical trial. We compared analytical performance across bDNA and two RT-qPCR workflows-RNA purification and a simplified NP-40 detergent-based treatment. A total of 77 clinical serum samples were analyzed. Method performance was assessed using assay precision, accuracy, and total error, along with linear regression and Bland-Altman analyses to evaluate inter-platform concordance. Noncompartmental PK analysis was performed on a subset of samples from four subjects. Results showed that RT-qPCR methods yielded lower mRNA concentrations than bDNA, with a consistent negative bias more pronounced in NP-40-treated samples. The purification RT-qPCR method showed closer agreement with bDNA across the quantitative range (R
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