A Cryoprotectant-Compatible Nanoporous Platform for Stable and Scalable Delivery of Biopharmaceuticals

CRISPR-Cas9 ribonucleoproteins (RNPs) represent a promising class of biopharmaceuticals for treating genetic and complex diseases. However, their clinical translation is limited by instability during storage and delivery. Lyophilization offers a potential solution, though conventional approaches often compromise structural integrity and bioactivity under non-cryogenic conditions. Here, we have developed a nanostructured delivery platform, designated Nano Banker & Blowball (NB²), which features a blowball-like architecture and tunable nanoscale pores. These pores are designed to protect RNPs and enable controlled release. The freeze-dried formulation (FNB²) integrates optimized cryoprotectants and a surface-engineered nanoparticle design, preserving morphology and function without excessive excipients. FNB² exhibits rapid rehydration and retains ∼70% of gene editing activity post-lyophilization, enabling robust functional gene editing in vitro and in vivo. It also maintains long-term stability and supports efficient cellular uptake, enabling administration via multiple routes. FNB² represents a scalable and robust platform for genetic therapeutics, vaccines, and biologics, particularly well-suited for resource-limited and emergency medical applications.