biostudies-literatureKilchrist KVNEI NIH HHSNIBIB NIH HHSNIDDK NIH HHSU.S. Department of Health and Human ServicesNHLBI NIH HHSU.S. Department of DefenseNCI NIH HHSVanderbilt UniversityNational Science Foundation1136-1152https://www.ebi.ac.uk/biostudies/studies/S-EPMC6995262biostudies-literatureUnknown13(2)Endolysosome entrapment is one of the key barriers to the therapeutic use of biologic drugs that act intracellularly. The screening of prospective nanoscale endosome-disrupting delivery technologies is currently limited by methods that are indirect and cumbersome. Here, we statistically validate Galectin 8 (Gal8) intracellular tracking as a superior approach that is direct, quantitative, and predictive of therapeutic cargo intracellular bioactivity through in vitro high-throughput screening and in vivo validation. Gal8 is a cytosolically dispersed protein that, when endosomes are disrupted, redistributes by binding to glycosylation moieties selectively located on the inner face of endosomal membranes. The quantitative redistribution of a Gal8 fluorescent fusion protein from the cytosol intACS nanoGal8 Visualization of Endosome Disruption Predicts Carrier-Mediated Biologic Drug Intracellular Bioavailability.PMC6995262R01CA224241U24 DK059637DGE-1445197P30 DK058404R01 EB019409DGE-0909667proposal number OR130302P30 EY008126R01 CA224241P30 DK020593R01EB019409W81XWH-14-1-0298P30 CA068485U2C DK059637R01 HL122347R01HL122347Kilchrist KVJackson MADuvall CLEvans BCBedingfield SKKelly IBWerfel TADailing EADimobi SCfalseGal8 Visualization of Endosome Disruption Predicts Carrier-Mediated Biologic Drug Intracellular Bioavailability.Endolysosome entrapment is one of the key barriers to the therapeutic use of biologic drugs that act intracellularly. The screening of prospective nanoscale endosome-disrupting delivery technologies is currently limited by methods that are indirect and cumbersome. Here, we statistically validate Galectin 8 (Gal8) intracellular tracking as a superior approach that is direct, quantitative, and predictive of therapeutic cargo intracellular bioactivity through in vitro high-throughput screening and in vivo validation. Gal8 is a cytosolically dispersed protein that, when endosomes are disrupted, redistributes by binding to glycosylation moieties selectively located on the inner face of endosomal membranes. The quantitative redistribution of a Gal8 fluorescent fusion protein from the cytosol int2019-01-01T00:00:00Z2019 Feb2025-04-19T09:34:32.5Z2020-05-22T10:50:06ZS-EPMC69952623062943110.1021/acsnano.8b05482