Nanoparticle Size Is a Critical Physicochemical Determinant of the Human Blood Plasma Corona: A Comprehensive Quantitative Proteomic Analysis

Abstract

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 particle's 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.

Effect of Silica Surface Properties on the Formation of Multilayer or Submonolayer Protein Hard Corona: Albumin Adsorption on Pyrolytic and Colloidal SiO2 Nanoparticles

“…57,58 Albumin is the most abundant component of the protein pool in FBS and FCS, as well in human blood. In one study, it was also found to be the most abundant protein in the hard corona of silica NPs, 59 while in other cases, its content in the hard protein corona was very low. [60][61][62] Physical parameters such as the surface curvature of NPs were proposed to play a role in the competitive adsorption of proteins in the process, resulting in the formation of the hard corona.…”

Synthesis, Functionalization, and Design of Magnetic Nanoparticles for Theranostic Applications

“…[675] As the field intensity decreases abruptly with distance, for targets that are more than a few centimeters deep in the body, the hydrodynamic force of blood flow overwhelms the magnetic force. [671] Thus, retention of the magnetic drug carrier particle (MDCPs) becomes quite low to the inherently weak nature of magnetic field, [7,14] which in turn, limits the location of this site to be less than a few centimeters deep inside the body. [660b] These obstacles in the way of performing external magnetic fields for MDT limit its application to superficial tumors and locations in the body that are very near to the surface of the skin wherein both strong magnetic field and field gradients as well as very low-velocity fields of blood flow are present.…”

“…Hence, a wide range of polymeric and biomimetic coatings as well as zwitterionic molecules have been shown promising to reduce the hydrophobicity of nanoparticles and inhibit the formation of protein corona. [14] In addition to evading the RES, cellular internalization of nanoparticles is also highly affected by their size and shape. Although larger nanoparticles increase the propensity of migration near the vascular wall, their chance to be taken up by tumor cells may be decreased.…”

In vitro biological effects of magnetic nanoparticles

“…Currently, several techniques are available for isolating nanoparticle-protein complexes, including equilibrium dialysis, size-exclusion chromatography, and microfiltration as being referred by Tenzer et al [16]. More recently, separation by centrifugation has been selected as common approach as it can easily be used routinely, be reproducible, and requires relatively little material [16,17].…”

“…More recently, separation by centrifugation has been selected as common approach as it can easily be used routinely, be reproducible, and requires relatively little material [16,17]. For the analysis of the composition of protein corona, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by liquid chromatography mass spectrometry (LC-MS) of digested peptides are used, combining with in silico analysis [16][17][18][19].…”