Multifunctional nanoparticle platform for targeted delivery and vaccines.

We describe a versatile, targeted delivery platform based on the Hepatitis B core protein virus-like particle (VLP). Multiple protein mutations were combined with cell-free protein synthesis and anaerobic processing to enable reliable production of nanoparticles (NPs) loaded with single or multiple cargoes (typically with concentration factors >10ˆ4) and functionalized with single or multiple surface adducts. Our design supports multiple functional requirements while also enabling flexible and reliable production. Process yields are about 6 x 10 NPs per mL of cell-free reaction; approximately 100-fold higher than current adeno-associated virus (AAV) yields and 8 times previously reported HBc VLP yields. We demonstrate platform feasibility and versatility by the surface display of a challenging-to-fold dengue fever antigen and by pharmacokinetic studies using whole-body mouse imaging. The platform supports rapid, parallel production of multiple product candidates to increase success rates for targeted therapeutics, gene therapies, imaging agents, and vaccines.