A Nanoparticle‐Integrated Complete Manufacturing Pipeline of Chemically Engineered Exosomes

Clinical translation of engineered exosomes, an emerging class of cell therapies, is hampered by challenges in each step of the manufacture flow, namely biogenesis, cargo loading, isolation, and storage. Here, we present a technology termed Tat-PNCAS-MIMS-MSC-Exo for manufacturing chemically engineered exosomes, with the above four steps being integrated by the use of the nanoparticle PNCAS-Tat (Tat peptide-conjugated protein-nanoparticle co-assembly supraparticle). This technology enables drastic improvements in all four steps of the manufacture flow of chemically engineered exosomes derived from mesenchymal stem cells (MSCs), a commonly-used cell type for cell therapies. The stimulation effect of exosome biogenesis by Tat peptide can be amplified by nanoparticle conjugation, a previously unknown nano-effect. The novel design of magnet setup MIMS (mobile internal magnetic separation) enables a unique capacity for scale-up of magnetic isolation, i.e., near-identical time for different scales to achieve near-complete isolation. This offers an effective solution to the long-standing problem of scale-up in applying magnetic isolation for biomanufacturing, which usually requires larger scales than bioanalytical applications. The manufacture process is robust, scalable, and economical. We conduct mechanistic studies of the nano-bio interactions, and demonstrate applications of the products in multiple disease models.