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Extracellular vesicles (EVs), including native red blood cell\u2010derived EVs (RBCEVs), offer biocompatibility but remain limited by heterogeneous composition, low yield, poor scalability, and limited control in cargo loading. Here, we introduce a bottom\u2010up strategy for developing engineered red blood cell extracellular vesicles (eRBCEVs) through microfluidic diffusional mixing of purified RBC lipids and components. Parametric optimization through multi\u2010physics simulation enabled predictive control over flow rates, lipid concentration, and channel geometry. eRBCEVs exhibited comparable encapsulation efficiency across molecular cargos of distinct sizes and properties, including oligonucleotides, gold nanoparticles, hemoglobin, erythrocruorin (\u223c3.6 MDa), and full adeno\u2010associated virus (AAV \u223c25\u00a0nm) particles, validated through cryo\u2010EM, chemical mapping, and high\u2010resolution TIRFM. Surface conjugation with CD47 peptide was established for comparison with RBCEVs; the bio\u2010conjugations with \u03b1\u2010PD\u2010L1 antibodies yielded functionalized vesicles with uptake in PD\u2010L1\u2010positive human tumor organoids. In vivo biodistribution and pharmacokinetic analysis in mice demonstrated sustained circulation and tissue distribution in the liver, spleen, and lungs. Neutrophil activation and macrophage uptake assays confirmed minimal immunogenicity compared to conventional particles and free\u2010protein controls. Collectively, our platform enables the scalable synthesis of customizable eRBCEVs that preserve RBC lipid features, support cargo flexibility, and potential for scale\u2010up. This work establishes eRBCEVs as a next\u2010generation therapeutic platform for programmable, patient\u2010specific delivery in gene therapy, immunomodulation, and translational nanomedicine.<\/jats:p>","DOI":"10.1002\/adhm.202504351","type":"journal-article","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T06:18:40Z","timestamp":1784269120000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Microfluidic Nano\u2010Assembly of Red\u2010Blood\u2010Cell (RBC) Lipids and Components for Engineering Extracellular Vesicles"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-8449-0724","authenticated-orcid":false,"given":"Chiranth K.","family":"Nagaraj","sequence":"first","affiliation":[{"name":"William G. 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