Physicochemical and cell biological aspects of microcondensate mediated intracellular infusion of antibodies

Physicochemical and cell biological aspects of microcondensate mediated intracellular infusion of antibodies | 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 Physicochemical and cell biological aspects of microcondensate mediated intracellular infusion of antibodies Shiroh Futaki, Yoshimasa Kawaguchi, Megumi Kiyokawa, Daisuke Yamasaki, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6563458/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Using engineered IgG antibodies with peptide tags consisting of glutamic acid and tyrosine residues, microcondensates were formed with the trimer of the attenuated cationic amphiphilic lytic peptide L17E conjugated to an Fc-binding peptide. Attachment of the microcondensates to the cell membrane resulted in immediate IgG infusion into the cells, regardless of the presence of serum. The unambiguous IgG and L17E structures allowed reliable evaluation of the physicochemical properties required for microcondensate formation and intracellular infusion, where the balanced stability of the microcondensates was found to be important. The infusion process was accompanied by a dynamic change in the cell membrane structure with filopodia-like membrane protrusions, trapping the microcondensates inside the cell. Inhibition of liquid-liquid phase separation (stress granule formation) of the intrinsically disordered and neurodegenerative disease-associated protein TDP-43 was also achieved using the IgG infusion system. Biological sciences/Drug discovery/Drug delivery Physical sciences/Materials science/Soft materials/Gels and hydrogels Biological sciences/Chemical biology/Protein design Biological sciences/Chemical biology/Mechanism of action Physical sciences/Materials science/Biomaterials/Drug delivery antibody microcondensate coacervate attenuated cationic amphiphilic lytic peptide intracellular delivery dynamic cytosolic infusion intrinsically disordered protein TDP-43 Full Text Additional Declarations Yes there is potential Competing Interest. Y.K., M.K. and S.F. are inventors on a patent application related to the technology described in this manuscript. All other authors declare no competing interests. Supplementary Files movie1.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-E20-E10(Alexa488part) and FcB(L17E)3 movie2.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-E20-(E4Y)2(Alexa488part) and FcB(L17E)3 movie3.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-E20-(EY)5(Alexa488part) and FcB(L17E)3 movie4.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-(E4Y)4-E10(Alexa488part) and FcB(L17E)3 movie5.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-(E4Y)4-(E4Y)2(Alexa488part) and FcB(L17E)3 movie6.mp4 FRAP imaging of microcondensates formed by LHAT-anti-GFP-IgG-(E4Y)4-(EY)5(Alexa488part) and FcB(L17E)3 movie7.mp4 Confocal timelapse imaging cells treated with microcondensates composed of LHAT-anti-GFP-IgG-E20-(E4Y)2(Alexa488part) and FcB(L17E)3 movie8.mp4 LLSM XY timelapse imaging of cells treated with microcondensates composed of LHAT-anti-GFP-IgG-E20-(E4Y)2(Alexa488part) and FcB(L17E)3 movie9.mp4 LLSM horizontal timelapse imaging of cells treated with microcondensates composed of LHAT-anti-GFP-IgG-E20-(E4Y)2(Alexa488part) and FcB(L17E)3 250429tagIgGSI.pdf Supporting Information reportingsummary.pdf Reporting Summary Cite Share Download PDF Status: Under Review Version 1 posted 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6563458","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":459356773,"identity":"21675879-7070-4b96-a84e-4adbd0752757","order_by":0,"name":"Shiroh 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