Protein Corona over Mesoporous Silica Nanoparticles: Influence of the Pore Diameter on Competitive Adsorption and Application to Prostate Cancer Diagnostics.

Diagnostic tests based on proteomics analysis can have significant advantages over more traditional biochemical tests. However, low molecular weight (MW) protein biomarkers are difficult to identify by standard mass spectrometric analysis, as they are usually present at low concentrations and are masked by more abundant resident proteins. We have previously shown that mesoporous silica nanoparticles are able to capture a predominantly low MW protein fraction from the serum, as compared to the protein corona (PC) adsorbed onto dense silica nanoparticles. In this study, we begin by further investigating this effect using liquid chromatography-mass spectrometry (LC-MS)/MS and thermogravimetric analysis (TGA) to compare the MW of the proteins in the coronas of mesoporous silica nanoparticles with the same particle size but different pore diameters. Next, we examine the process by which two proteins, one small and one large, adsorb onto these mesoporous silica nanoparticles to establish a theory of why the corona becomes enriched in low MW proteins. Finally, we use this information to develop a novel system for the diagnosis of prostate cancer. An elastic net statistical model was applied to LC-MS/MS protein coronas from the serum of 22 cancer patients, identifying proteins specific to each patient group. These studies help to explain why low MW proteins predominate in the coronas of mesoporous silica nanoparticles, and they illustrate the ability of this information to supplement more traditional diagnostic tests.

Spatial Mapping of Protein Adsorption on Mesoporous Silica Nanoparticles by Stochastic Optical Reconstruction Microscopy.

Exposure to biological fluid envelops a nanoparticle in layers of proteins and biomolecules, which has a profound impact on the nanoparticle's biological fate. Although the identities and amounts of the proteins in this "corona" have been thoroughly examined, the spatial arrangement of the proteins is unclear, a problem that is compounded on porous nanoparticles due to penetration of proteins within the porous network. To address this problem, we have developed a procedure based on information derived from stochastic optical reconstruction microscopy. We employed a mathematical model to reveal the penetration depth of several proteins within porous nanoparticles. Understanding protein penetration depth provides an explanation for the composition of the protein corona, aiding in the development of safe and effective particle-based therapies.

Protein Adsorption From Biofluids on Silica Nanoparticles: Corona Analysis as a Function of Particle Diameter and Porosity.

A study on the adsorption of proteins from fetal bovine serum (FBS) on spherical dense and mesoporous silica nanoparticles with a wide range of diameters, from 70 to 900 nm, is presented. Monodisperse populations of particles with a range of diameters were obtained through modifications of the Stöber method. Extensive characterization of the particles was then performed using N2 physisorption, TEM, DLS, and ζ-potential. Following serum exposure, proteomic evaluation in concert with thermogravimetric analysis revealed the associated concentrations of each protein identified in the hard corona. Small particles adsorbed the largest amount of protein, due to their larger external surface area. Proteins with low molecular weights (<50 kDa) constituted the majority of the protein corona, totaling between 60 and 80% of the total mass of adsorbed protein. Here, the higher surface curvature of small particles favors the enrichment of smaller proteins. Porosity does not promote protein adsorption but improves deposition of the low molecular weight protein fraction due to the size-exclusion effect related to pore diameter. These results have important implications for the use of dense and porous silica nanoparticles in biomedical applications.