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In biological fluids, proteins associate with nanoparticles, leading to a protein 'corona' defining the biological identity of the particle. However, a comprehensive knowledge of particle-guided protein fingerprints and their dependence on nanomaterial properties is incomplete. We studied the long-lived ('hard') blood plasma derived corona on monodispersed amorphous silica nanoparticles differing in size (20, 30, and 100 nm). Employing label-free liquid chromatography mass spectrometry, one- and two-dimensional gel electrophoresis, and immunoblotting the composition of the protein corona was analyzed not only qualitatively but also quantitatively. Detected proteins were bioinformatically classified according to their physicochemical and biological properties. Binding of the 125 identified proteins did not simply reflect their relative abundance in the plasma but revealed an enrichment of specific lipoproteins as well as proteins involved in coagulation and the complement pathway. In contrast, immunoglobulins and acute phase response proteins displayed a lower affinity for the particles. Protein decoration of the negatively charged particles did not correlate with protein size or charge, demonstrating that electrostatic effects alone are not the major driving force regulating the nanoparticle–protein interaction. Remarkably, even differences in particle size of only 10 nm significantly determined the nanoparticle corona, although no clear correlation with particle surface volume, protein size, or charge was evident. Particle size quantitatively influenced the particles decoration with 37% of all identified proteins, including (patho)biologically relevant candidates. We demonstrate the complexity of the plasma corona and its still unresolved physicochemical regulation, which need to be considered in nanobioscience in the future.
Deliberate and accidental exposure of the ecosystem including humans to nanoparticles becomes inevitable as nanomaterials are increasingly used. In biological fluids, biomolecules associate with nanoparticles, leading to the formation of a dynamic biomolecule “corona” that critically defines the biological identity of the particle. As the bio-physical properties of such a decorated particle often differ significantly from those of the formulated particle a detailed characterization of nanoparticles in biological environments becomes increasingly important though, nevertheless also technically challenging. Here, we introduce experimental methods currently employed for nanoparticle characterization, present examples underlining the complexity of the nano-bio interface, and discuss the need for further technical and conceptual developments. A deep and mechanistic bio-physical understanding of the nano-bio interface is a challenge but also fundamental prerequisite for future applications in nanobiology, nanomedicine and nano(eco)toxicology.