{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T06:57:19Z","timestamp":1790578639831,"version":"4.1.0"},"reference-count":23,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["advanced.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Advanced Materials"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Lipid nanoparticles (LNPs) are the leading platform for delivery of RNA\u2010based therapeutics. There is considerable interest in optimizing the transfection potency of these systems. Standard formulation protocols involve mixing lipids in ethanol with a buffer containing mRNA at pH 4, followed by dialysis against a pH 7.4 saline buffer to remove ethanol and raise the pH. The dialysis process results in fusion of mRNA\u2010lipid complexes and small vesicles formed at pH 4, driven by two processes: Conversion of the ionizable lipid from the positively charged bilayer form to the neutral oil form and fusion stimulated by higher ionic strengths. Here we explored the consequences of a two\u2010step dialysis process involving formulating at pH 4, 25\u00a0mM NaOAc, and then inducing fusion at pH 4 by dialyzing against a higher ionic strength buffer while maintaining the ethanol concentration at 25% (v\/v). This was followed by dialysis against PBS to raise the pH. We show that the two\u2010step protocol results in increased mRNA loading per LNP, a decreased proportion of \u201cempties\u201d as compared to the standard process, and improved transfection potencies in vitro and in vivo when compared to LNP mRNA formed from the traditional one\u2010step buffer exchange process.<\/jats:p>","DOI":"10.1002\/adma.75174","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T06:35:59Z","timestamp":1790577359000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Increasing Lipid Nanoparticle mRNA Copy Number Improves Transfection Potency"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2783-1317","authenticated-orcid":false,"given":"Yao","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"School of Biomedical Engineering University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8714-8440","authenticated-orcid":false,"given":"Jerry","family":"Leung","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6571-021X","authenticated-orcid":false,"given":"Judy","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Psychiatry Djavad Mowafaghian Centre for Brain Health University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1614-4079","authenticated-orcid":false,"given":"Arash","family":"Momeni","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6790-1140","authenticated-orcid":false,"given":"Albert","family":"Kamanzi","sequence":"additional","affiliation":[{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ariadne Tuckmantel","family":"Bido","sequence":"additional","affiliation":[{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suiyang","family":"Liao","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"School of Biomedical Engineering University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Kang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4031-9932","authenticated-orcid":false,"given":"Colton","family":"Strong","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3644-0019","authenticated-orcid":false,"given":"Christian J.","family":"Kastrup","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"},{"name":"Department of Surgery Medical College of Wisconsin  Milwaukee Wisconsin USA"},{"name":"Versiti Blood Research Institute  Milwaukee Wisconsin USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Leslie","sequence":"additional","affiliation":[{"name":"Michael Smith Laboratories University of British Columbia  Vancouver British Columbia Canada"},{"name":"Department of Physics and Astronomy University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0261-4642","authenticated-orcid":false,"given":"Miffy Hok Yan","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia Canada"},{"name":"Faculty of Pharmaceutical Sciences University of British Columbia  Vancouver British Columbia Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pieter R.","family":"Cullis","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of British Columbia  Vancouver British Columbia 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