Isolation and Quantification of miRNA from the Biomolecular Corona on Mesoporous Silica Nanoparticles

Abstract

To understand the factors that control the formation of the biomolecular corona, a systematic study of the adsorption of several miRNAs shown to be important in prostate cancer on amine-functionalized mesoporous silica nanoparticles (MSN-NH2) has been performed. Process parameters including miRNA type, nanoparticle concentration, incubation temperature and incubation time were investigated, as well as the potential competition for adsorption between different miRNA molecules. The influence of proteins and particle PEGylation on miRNA adsorption were also explored. We found that low particle concentrations and physiological temperature both led to increased miRNA adsorption. Adsorption of miRNA was also higher when proteins were present in the same solution; reducing or preventing protein adsorption by PEGylating the MSNs hindered adsorption. Finally, the amount of miRNA adsorbed from human serum by MSN-NH2 was compared to a commercial miRNA purification kit (TaqMan®, Life Technologies, Carlsbad, CA, USA). MSN-NH2 adsorbed six times as much miRNA as the commercial kit, demonstrating higher sensitivity to subtle up- and downregulation of circulating miRNA in the blood of patients.

Influence of surface chemistry and morphology of nanoparticles on protein corona formation

“…In the last few years, the adsorption of a wider range of biomolecules on the surface of NPs has been studied. By coupling chromatographic methods and mass spectrometry, nanomaterials have been shown to adsorb small molecules such as lipids (Gunnarsson et al, 2018; La Barbera et al, 2020; Lee et al, 2018; Lima et al, 2020), amino acids (Pink et al, 2018) and oligonucleotides (Vidaurre‐Agut et al, 2021), with lipids being nowadays the most investigated. In a recent study, Chetwynd et al demonstrated that, similarly to PC, the corona of metabolites can vary significantly between silica, TiO 2 , and polystyrene NPs, indicating a possible role of surface chemistry of materials on their interactions with small molecules (Chetwynd et al, 2020).…”

Mechanistic Understanding of Protein Corona Formation around Nanoparticles: Old Puzzles and New Insights

“…Studies of in vivo‐formed coronas have recently appeared as well. [ 201,209–212 ] In this setting, other types of biomolecules including nucleic acids, [ 213 ] lipids, [ 214,215 ] sugars, [ 23 ] and small metabolites [ 216 ] are present and adsorb to the NPs, and take part in exchange processes that continuously modify the composition of the biomolecular corona. In their comprehensive LC‐MS/MS proteomic analysis, Hadjidemetriou et al.…”

“…Studies of in vivo-formed coronas have recently appeared as well. [201,[209][210][211][212] In this setting, other types of biomolecules including nucleic acids, [213] lipids, [214,215] sugars, [23] and small metabolites [216] are present and adsorb to the NPs, and take part in exchange processes that continuously modify the composition of the biomolecular corona. In their comprehensive LC-MS/MS proteomic analysis, Hadjidemetriou et al [209] observed significant differences in protein composition in the coronas formed on PEGylated liposomes (with and without attached antibodies for targeting) in vitro (10 min incubation in CD-1 mouse plasma) and in vivo (NP recovery from CD-1 mice 10 min after intravenous injection).…”