In vivo formation of protein corona on gold nanoparticles. The effect of their size and shape

Abstract

The efficacy of drug delivery and other nanomedicine-related therapies largely relies on the ability of nanoparticles to reach the target organ. However, when nanoparticles are injected into the bloodstream, their surface is instantly modified upon interaction with blood components, principally with proteins. It is well known that a dynamic and multi-layered protein structure is formed spontaneously on the nanoparticle upon contact with physiological media, which has been termed protein corona. Although several determinant factors involved in protein corona formation have been identified from in vitro studies, specific relationships between the nanomaterial synthetic identity and its ensuing biological identity under realistic in vivo conditions remain elusive. We present here a detailed study of in vivo protein corona formation after blood circulation of anisotropic gold nanoparticles (nanorods and nanostars). Plasmonic gold nanoparticles of different shapes and sizes were coated with polyethyleneglycol, intravenously administered in CD-1 mice and subsequently recovered. The results from gel electrophoresis and mass spectrometry analysis revealed the formation of complex protein coronas, as early as 10 minutes post-injection. The total amount of protein adsorbed onto the particle surface and the protein corona composition were found to be affected by both the particle size and shape.

Biological Behavior Regulation of Gold Nanoparticles via the Protein Corona

“…Using the methodology based on a combination of size exclusion chromatography and membrane ultrafiltration, García‐Álvarez et al first reported in vivo PC formation on polyethylene glycol (PEG)‐coated gold nanorods (GNRs) and gold nanostars (GNSs). [ 34 ] They found that the first and second highest proteins in the PC formed in blood were serum albumin and alpha‐2‐macroglobulin (A2M), respectively, and the PC composition was size‐ and shape‐dependent. However, due to the difficulties in the in vivo sample pretreatment, not all of the injected GNPs were recovered, and some proteins may not detach from the NP surface for analysis.…”

“…A comparative study of the PC formation on GNRs [ 43 ] and GNSs [ 44 ] in vivo with different sizes showed that the total amount of protein adsorbed on gold nanomaterials was affected by both size and shape of NPs. [ 34 ] The larger GNRs and GNSs (70 nm) adsorbed larger amounts of proteins than smaller GNRs and GNSs (40 nm). Additionally, the amounts of proteins covering on the surface of GNSs were significantly larger than those of GNRs with the same size due to the higher surface area of GNSs.…”

“…Although it is now accepted that the proteins in the PC could modulate the accumulation of NPs in different organs and tissues, the difficulty in recovering the GNPs from blood and all of the adsorbed proteins from the GNP surface for analysis [ 34 ] limited the feasibility to predict the biodistribution of GNPs in vivo. So far, the in vivo biodistribution of GNPs has only been defined in the case of a few corona proteins used as pre‐coated proteins.…”

Functionalized gold nanoparticles for sample preparation: A review

“…The main principle of indirect quantitation of the protein surface coverage is to measure the protein concentration before and after the conjugation in the supernatant and based on mass difference, the conjugated amount of protein can be backcalculated. Common methods for protein quantification are Lowry assay , Bradford assay , and bicinchoninic acid assay . They are the most popular methods for this purpose and allow sensitive detection of free proteins in the reaction mixture after removal of the protein GNP bioconjugates.…”

Star‐Shaped Gold Nanoparticles as Friendly Interfaces for Protein Electrochemistry: the Case Study of Cytochrome c

“…For example, adsorption of proteins on AuNPs of different sizes can lead to different degrees of structural changes, due to the variation of curvature with size in spherical nanoparticles . Shape can also influence adsorption and reactivity, because non‐spherical nanoparticles expose high index crystal facets that have higher binding energies with the molecules adsorbed. Surface chemistry influences not only the extent of protein adsorption, but also the degree of protein denaturation upon adsorption.…”