Multivalent Display Using Hybrid Virus Nanoparticles
Many important biological interactions are multivalent and often sensitive to spatial organization. Nonenveloped viruses are a natural source of scaffolds for building multivalent ligands to probe these types of interactions which avoid complex synthetic schemes required for other types of scaffolds. The coat protein (CP) of bacteriophage Qβ can be fused to protein domains and coexpressed with the unfused CP to produce hybrid nanoparticles with high exterior loading of xenogenic protein domains. These hybrid nanoparticles are simple to produce in large quantity. Starting from cDNAs for the virus CP and a codon-optimized ligand domain of interest, bulk purification can be completed in as little as 3 weeks. Major phases of the work involve the cloning of cDNAs into plasmid vectors, test expressions for hybrid nanoparticle formation, and purification by selective precipitation and ultracentrifugation. For uncomplicated protein domains, laboratory culture yields as high as 50 mg/L and 30 protein domains per particle have been routinely achieved.
Key wordsNanoparticle Protein expression Protein purification Multivalency Gene engineering
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