Scientific Publications
Technical tip: high-resolution isolation of nanoparticle–protein corona complexes from physiological fluids
Nanoscale, 7, 11980-11990
Nanoparticles (NPs) in contact with biological fluids are generally coated with environmental proteins,forming a stronger layer of proteins around the NP surface called the hard corona. Protein corona complexesprovide the biological identity of the NPs and their isolation and characterization are essential tounderstand their in vitro and in vivo behaviour. Here we present a one-step methodology to recover NPsfrom complex biological media in a stable non-aggregated form without affecting the structure or compositionof the corona. This method allows NPs to be separated from complex fluids containing biologicalparticulates and in a form suitable for use in further experiments. The study has been performed systematicallycomparing the new proposed methodology to standard approaches for a wide panel of NPs. NPswere first incubated in the biological fluid and successively recovered by sucrose gradient ultracentrifugationin order to separate the NPs and their protein corona from the loosely bound proteins. The isolatedNP–protein complexes were characterized by size and protein composition through Dynamic Light Scattering,Nanoparticle Tracking Analysis, SDS-PAGE and LC-MS. The protocol described is versatile and canbe applied to diverse nanomaterials and complex fluids. It is shown to have higher resolution in separatingthe multiple protein corona complexes from a biological environment with a much lower impact on theirin situ structure compared to conventional centrifugal approaches.
Nanoscale, 7, 11980-11990
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