{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T22:28:07Z","timestamp":1791930487196,"version":"4.3.4"},"reference-count":39,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"EU Joint Research Centre projects \u201cInnovative Biotechnologies in a Competitive Europe\u201d","award":["34092\u2014INNOBIOTECH"],"award-info":[{"award-number":["34092\u2014INNOBIOTECH"]}]},{"name":"Innovative Tests and Standards for EU Pathogens Surveillance","award":["32181\u2014iTEST"],"award-info":[{"award-number":["32181\u2014iTEST"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>The approval of safe and effective lipid nanoparticles-encapsulated mRNA (LNP-mRNA) vaccines during the SARS-CoV-2 pandemic is catalysing the development of the next generation of mRNA therapeutics. Accurate characterisation methods are crucial for assessing the quality and efficacy of these complex formulations. Several analytical techniques exist for the quantification of the mRNA drug substance, including UV spectroscopy, capillary electrophoresis, high-performance liquid chromatography (HPLC), and digital PCR (dPCR). Here, we compare these methods, highlight their limitations, and discuss critical considerations for experimental design to improve the agreement between different analytical methods.<\/jats:p>","DOI":"10.3390\/ijms27167379","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:09:32Z","timestamp":1787054972000},"page":"7379","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Critical Evaluation of Key Methods for RNA Quantification"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4847-9992","authenticated-orcid":false,"given":"Valentina","family":"Paracchini","sequence":"first","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 2440 Geel, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6782-4704","authenticated-orcid":false,"given":"Mauro","family":"Petrillo","sequence":"additional","affiliation":[{"name":"Seidor Italy SRL, 20129 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Fornara","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7150-9523","authenticated-orcid":false,"given":"Sabrina","family":"Gioria","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4899-5284","authenticated-orcid":false,"given":"Gabriele","family":"Leoni","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1446-0556","authenticated-orcid":false,"given":"Giuditta","family":"Guerrini","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5371-3798","authenticated-orcid":false,"given":"Otmar","family":"Geiss","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3157-4552","authenticated-orcid":false,"given":"Antonio","family":"Marchini","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8474-7974","authenticated-orcid":false,"given":"Luigi","family":"Calzolai","sequence":"additional","affiliation":[{"name":"European Commission, Joint Research Centre (JRC), 21027 Ispra, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2702","DOI":"10.1016\/j.ymthe.2023.07.022","article-title":"Development of an mRNA-lipid nanoparticle vaccine against Lyme disease","volume":"31","author":"Pine","year":"2023","journal-title":"Mol. 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Data"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7379\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:09:52Z","timestamp":1787054992000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7379"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,18]]},"references-count":39,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27167379"],"URL":"https:\/\/doi.org\/10.3390\/ijms27167379","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,18]]}}}