High Resolution ES-DMA quantifies AAV capsid DNA content by electrical mobility to mass correlation

High Resolution ES-DMA quantifies AAV capsid DNA content by electrical mobility to mass correlation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article High Resolution ES-DMA quantifies AAV capsid DNA content by electrical mobility to mass correlation Luis Javier Perez Lorenzo, Preston Dennett, Lohra Young, Benjamin Draper, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8291101/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 10 You are reading this latest preprint version Abstract Recombinant Adeno-Associated Virus (rAAV) is the leading viral vector platform for gene therapy. A persistent challenge in rAAV manufacturing and quality control is accurate assessment of the DNA content of purified viral capsids. Empty particles, as well as ‘partial’ and over-filled species commonly contaminate even highly purified rAAV preparations. These impurities contribute to high production costs and reduced therapeutic efficacy, resulting in higher required doses and increased risks to patient health. Here we present high-resolution electrospray differential mobility analysis (ES-DMA) as a novel benchtop analytical modality for rAAV vector characterization. Sampling microliter volumes of analyte, this method achieves angstrom-scale particle sizing which we demonstrate is sufficient to resolve DNA encapsidation by rAAVs via differential electrical mobility. We benchmark this approach against charge detection mass spectrometry (CD-MS), an emerging gold standard for rAAV analytics, and identify a robust near-linear correlation between electrical mobility and mass of aerosolized rAAV particles. This relationship enables rapid identification and relative quantification of empty, full, and partial / over-filled rAAV capsids by ES-DMA. Biological sciences/Biological techniques/Biophysical methods Biological sciences/Biotechnology/Gene delivery/Genetic vectors Full Text Additional Declarations Yes there is potential conflict of interest. P.D., J.S. and L.J.P-L are employees of NanoEngineering Corporation, which is involved in the commercialization of the ES-DMA instrument used in this study. L.M.Y, B.E.D. and M. J. are employees of Megadalton Solutions which has developed CD-MS instrumentation technology (licensed to Waters Corporation for commercialization), and currently offers commercial rAAV testing services. Supplementary Files Tablesi2.xlsx Table SI2 Supplementalinformationfinal.pdf Supplemental Information: Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: revise 06 Feb, 2026 Review # 3 received at journal 27 Jan, 2026 Review # 1 received at journal 29 Dec, 2025 Reviewer # 3 agreed at journal 19 Dec, 2025 Reviewer # 2 agreed at journal 12 Dec, 2025 Reviewer # 1 agreed at journal 12 Dec, 2025 Reviewers invited by journal 11 Dec, 2025 Editor assigned by journal 10 Dec, 2025 Submission checks completed at journal 10 Dec, 2025 First submitted to journal 05 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8291101","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":559063348,"identity":"806725e1-5b25-48dc-b149-822ba0546723","order_by":0,"name":"Luis Javier Perez 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