{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T11:25:57Z","timestamp":1783596357804,"version":"3.55.0"},"reference-count":18,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100014726","name":"National Institute for Innovation in Manufacturing Biopharmaceuticals","doi-asserted-by":"publisher","award":["NANB17H002"],"award-info":[{"award-number":["NANB17H002"]}],"id":[{"id":"10.13039\/100014726","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Gates Foundation","doi-asserted-by":"publisher","award":["INV-046943"],"award-info":[{"award-number":["INV-046943"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["gatesopenresearch.org"],"crossmark-restriction":false},"short-container-title":["GatesOpenRes"],"abstract":"<jats:p>Potency assays assess the functional integrity of a target antigen to ensure that the final drug product is consistent between the commercial and clinical trial lots. Existing mRNA-LNP vaccine potency assays require complex quantification methods, such as flow cytometry, which are not widely adaptable to routine use by low- and middle-income country (LMIC) manufacturers or National Regulatory Agency (NRA) laboratories. To address the need for more accessible mRNA-LNP vaccine potency methods, an enzyme-linked immunosorbent assay (ELISA) was developed to quantify the target proteins produced by an mRNA-LNP vaccine after cell transfection. SARS-CoV-2 mRNA-LNP targeting the wild-type (Wuhan) strain was used as a model vaccine for the development and qualification of an ELISA method. The adaptability of the ELISA to other mRNA-LNP vaccines was demonstrated by altering the method used for quantification of a commercial Omicron mRNA-LNP vaccine. Alternative mRNA-LNP potency methods, such as western blotting, fluorescence focus assay (FFA), and direct ELISA were used as analogous methods for comparison. The developed quantitative stability-indicating ELISA is a globally accessible potency method that utilizes existing equipment and knowledge. Testing conducted on wild type SARS CoV-2 and Omicron vaccines demonstrates the adaptability of the method to other mRNA vaccine targets.<\/jats:p>","DOI":"10.12688\/gatesopenres.16382.1","type":"journal-article","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T10:49:08Z","timestamp":1783594148000},"update-policy":"https:\/\/doi.org\/10.12688\/gatesopenres.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Qualification of a quantitative stability indicating potency assay for mRNA-LNP vaccine candidates"],"prefix":"10.12688","author":[{"given":"Anan","family":"Bzami","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcus","family":"Estrada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tim","family":"Schofield","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica A","family":"White","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2026]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"261-279","DOI":"10.1038\/nrd.2017.243","article-title":"mRNA vaccines - a new era in vaccinology.","volume":"17","author":"N Pardi","year":"2018","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1465-1468","DOI":"10.1126\/science.1690918","article-title":"Direct gene transfer into mouse muscle in vivo.","volume":"247","author":"J Wolff","year":"1990","journal-title":"Science."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1038\/s41541-022-00470-4","article-title":"Development of functionally relevant potency assays for monovalent and multivalent vaccines delivered by evolving technologies.","volume":"7","author":"G Sanyal","year":"2022","journal-title":"NPJ Vaccines."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1038\/s41541-021-00317-4","article-title":"Considerations for bioanalytical characterization and batch release of COVID-19 vaccines.","volume":"6","author":"G Sanyal","year":"2021","journal-title":"NPJ Vaccines."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"163-180","DOI":"10.4155\/bio-2017-0176","article-title":"Principles of vaccine potency assays.","volume":"10","author":"T Verch","year":"2018","journal-title":"Bioanalysis."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1704","DOI":"10.3390\/vaccines10101704","article-title":"Rapid Identity and Quantity CQA Test for Multivalent mRNA Drug Product Formulations.","volume":"10","author":"R Gao","year":"2022","journal-title":"Vaccines."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1144","DOI":"10.3390\/vaccines12101144","article-title":"Analytical Performance of a Multiplexed Microarray Assay for Rapid Identification and Quantification of a Multivalent mRNA Vaccine.","volume":"12","author":"M Gerold","year":"2024","journal-title":"Vaccines."},{"key":"ref8","article-title":"USP_mRNA_Vaccines_One_Pager_v5_20240618_FINAL 1."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"594","DOI":"10.3389\/fimmu.2019.00594","article-title":"Advances in mRNA Vaccines for Infectious Diseases.","volume":"10","author":"C Zhang","year":"2019","journal-title":"Front. Immunol."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"283-289","DOI":"10.1038\/s41586-021-03275-y","article-title":"BNT162b vaccines protect rhesus macaques from SARS-CoV-2.","volume":"592","author":"A Vogel","year":"2021","journal-title":"Nature."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"19579","DOI":"10.1038\/s41598-021-98972-z","article-title":"Viral detection and identification in 20 min by rapid single-particle fluorescence in-situ hybridization of viral RNA.","volume":"11","author":"C Hepp","year":"2021","journal-title":"Sci. Rep."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"e0004746","DOI":"10.1371\/journal.pntd.0004746","article-title":"An mRNA Vaccine Encoding Rabies Virus Glycoprotein Induces Protection against Lethal Infection in Mice and Correlates of Protection in Adult and Newborn Pigs.","volume":"10","author":"M Schnee","year":"2016","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"e0006108","DOI":"10.1371\/journal.pntd.0006108","article-title":"A thermostable messenger RNA based vaccine against rabies.","volume":"11","author":"L Stitz","year":"2017","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref14","doi-asserted-by":"publisher","article-title":"<1033> Biological Assay Validation.","DOI":"10.31003\/USPNF_M912_01_01"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"e0260173","DOI":"10.1371\/journal.pone.0260173","article-title":"Direct addition of poly-lysine or poly-ethylenimine to the medium: A simple alternative to plate pre-coating.","volume":"17","author":"A Faussner","year":"2022","journal-title":"PloS One."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"115523","DOI":"10.1016\/j.jpba.2023.115523","article-title":"Development and qualification of cell-based relative potency assay for a human respiratory syncytial virus (RSV) mRNA vaccine.","volume":"234","author":"H Li","year":"2023","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"1364-1371","DOI":"10.1016\/j.xphs.2023.01.007","article-title":"Characterization of BNT162b2 mRNA to Evaluate Risk of Off-Target Antigen Translation.","volume":"112","author":"H Patel","year":"2023","journal-title":"J. Pharm. Sci."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"6709-6722","DOI":"10.1021\/acsnano.0c10064","article-title":"Apolipoprotein E Binding Drives Structural and Compositional Rearrangement of mRNA-Containing Lipid Nanoparticles.","volume":"15","author":"F Sebastiani","year":"2021","journal-title":"ACS Nano."}],"container-title":["Gates Open Research"],"original-title":[],"link":[{"URL":"https:\/\/gatesopenresearch.org\/articles\/10-26\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/gatesopenresearch.org\/articles\/10-26\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/gatesopenresearch.org\/articles\/10-26\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T11:06:41Z","timestamp":1783595201000},"score":1,"resource":{"primary":{"URL":"https:\/\/gatesopenresearch.org\/articles\/10-26\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026]]},"references-count":18,"URL":"https:\/\/doi.org\/10.12688\/gatesopenres.16382.1","relation":{},"ISSN":["2572-4754"],"issn-type":[{"value":"2572-4754","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026]]},"assertion":[{"value":"Awaiting Peer Review","URL":"https:\/\/gatesopenresearch.org\/articles\/10-26\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"This work was performed under a Project Award Agreement from the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) and a financial assistance award 70NANB17H002 from the U.S. Department of Commerce, National Institute of Standards and Technology. This study was partially supported by the Gates Foundation [INV-046943]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":0,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}