In Vitro Assembly of Virus-Derived Designer Shells Around Inorganic Nanoparticles

Abstract

Nanoparticle-templated assembly of virus shells provides a promising approach to the production of hybrid nanomaterials and a potential avenue toward new mechanistic insights in virus phenomena originating in many-body effects, which cannot be understood from examining the properties of molecular subunits alone. This approach complements the successful molecular biology perspective traditionally used in virology, and promises a deeper understanding of viruses and virus-like particles through an expanded methodological toolbox. Here we present protocols for forming a virus coat protein shell around functionalized inorganic nanoparticles.

Hydrophobic Cargo Encapsulation into Virus Protein Cages by Self-Assembly in an Aprotic Organic Solvent

“…BMV was purified from infected Atlantic barley (Hordeum vulgare) leaves, 71 and capsid proteins were purified as previously described. 36 BMV virions were disassembled by 24 h dialysis against disassembly buffer (0. SDS-PAGE was used to ascertain the integrity of the purified protein.…”

“…VLP Assembly (Around AuNP and UCNP). VLPs were synthesized as reported previously 36 with slight modifications in buffers and protein to nanoparticle ratio. 260 BMV capsid proteins (CPs) were used per AuNP-12.3 and 870 CP for the encapsulation of AuNP-25.8, UCNP-24.2, and UCNP-28.6.…”

“…35 Usually, in vitro synthesis of VLPs is carried out in aqueous media. 36 To the best of our knowledge, self-assembly of VLPs in DMSO-containing solutions has never been studied. Presumably, the known DMSO ability to inactivate viruses and denature certain proteins 26,37,38 has dampened the enthusiasm for such explorations.…”

“…Usually, in vitro synthesis of VLPs is carried out in aqueous media . To the best of our knowledge, self-assembly of VLPs in DMSO-containing solutions has never been studied.…”

Harnessing physicochemical properties of virus capsids for designing enzyme confined nanocompartments

“…Co-expression of AaLS-neg and GFP-R10 in Escherichia coli led to the formation of an AaLS-neg cage loaded with GFP-R10 ( Figure 2a ) [ 34 ]. Electrostatic interactions between CPs and cargos have also been applied to encapsulate non-proteinaceous cargos, such as pre-formed inorganic nanoparticles inside of VLPs derived from several different viruses [ 36 - 38 ].…”