End-to-End Biomanufacturing of Extracellular Vesicles

Extracellular vesicles (EVs) have emerged as next-generation, multispecific, and multifunctional agents across medicine, biotechnology, veterinary science, cosmetics, and nutrition. Their delivery efficiency of therapeutic cargo surpasses that of conventional carriers, and their capacity to simultaneously transport internal cargo and activate multiple recipient cells gives them a therapeutic versatility that no other platform can match. Translating this potential into real-world products depends on integrating process design with product characterization, a link that has not yet been systematically established for EV manufacturing. The functionality of an EV is inseparable from how it is made. EV composition is shaped at every stage of production, from the biology of the producer cell, to upstream microenvironmental stressors, to the operational parameters of downstream isolation and purification. Each of these variables influences the final product quality and functionality, which means that process design is, in effect, product design. While many guidelines, including minimal information for studies of extracellular vesicles (MISEV) and quality by design (QbD), have been reported, they operate at different levels: MISEV ensures the validity of EV characterization, whereas QbD defines how a manufacturer designs, controls, and releases a product. Currently, however, there is no link between them to provide an integrative framework. This manuscript, therefore reviews and proposes a framework for context-of-use–dependent EV biomanufacturing, in which biomanufacturing and bioprocessing parameters are deliberately matched to the intended application. By linking specific process conditions to resultant EV characteristics, and by fine-tuning upstream and downstream operations accordingly, bespoke EVs can be produced for defined therapeutic, diagnostic, or industrial purposes rather than made as a single generic product. Together, this work offers essential guidance for producing EVs at scale and for designing end-to-end bioprocesses that can be tailored to the diverse needs of future EV-based products.