Approach For Efficient Protein Incorporation Into Recombinant Vaults
SUMMARY
UCLA researchers in the departments of Medicine, Microbiology, Immunology & Molecular Genetics, and Bioengineering have developed a novel method for loading protein payloads into vault nanoparticle carriers.
BACKGROUND
Vaults are naturally occurring nanoparticles that are widely present in eukaryotes. There is significant interest in developing these materials as delivery vehicles for small molecules, proteins, or antigens as vaccine vectors. Methods currently in use for loading biomolecules include modification of the protein of interest with a vault-binding domain, or direct genetic fusion to the major vault protein itself. However, these methods are inconsistent, require lengthy optimization processes, or interfere with vault assembly. Developing methodology to load protein payloads into vaults will therefore be critically important to their use as protein delivery vehicles.
INNOVATION
Prof. Yang and coworkers have developed a novel method for loading protein payloads into vault nanoparticle carriers. A specific region in the vault structure has been identified where the amino acids are disordered. Because this region is flexible, the area can be tolerant of protein insertions. Proof of concept has been demonstrated by genetic insertion of proteins into the disordered loop with subsequent incorporation of desired sequences. No disruption of normal vault morphology was observed.
APPLICATIONS
- Delivery of antigens for vaccines
- Delivery of functional proteins
- Delivery of small molecules via protein carriers
ADVANTAGES
- Modification in flexible regime does not interfere with vault assembly
- Generalizable to multiple types of proteins
- More reliable than current state of the art techniques
- No protein-specific optimization required
STATE OF DEVELOPMENT
Proof of concept has been demonstrated by inserting a restriction enzyme site into the disordered loop.