Protein Corona of Nanoparticles: Distinct Proteins Regulate the Cellular Uptake

Abstract

Understanding nanoparticle-protein interactions is a crucial issue in the development of targeted nanomaterial delivery. Besides unraveling the composition of the nanoparticle's protein coronas, distinct proteins thereof could control nanoparticle uptake into specific cell types. Here we differentially analyzed the protein corona composition on four polymeric differently functionalized nanoparticles by label-free quantitative mass spectrometry. Next, we correlated the relative abundance of identified proteins in the corona with enhanced or decreased cellular uptake of nanoparticles into human cancer and bone marrow stem cells to identify key candidates. Finally, we verified these candidate proteins by artificially decorating nanoparticles with individual proteins showing that nanoparticles precoated with the apolipoproteins ApoA4 or ApoC3 significantly decreased the cellular uptake, whereas precoating with ApoH increased the cellular uptake.

The Impact of Nanoparticle Protein Corona on Cytotoxicity, Immunotoxicity and Target Drug Delivery

“…Among the latters, label-free MS [68] is becoming the gold standard procedure with which to obtain simultaneously a quantitative and qualitative analysis of PC components [64,[69][70][71] thanks to its benefits that include a low number of steps in the protocol and high reproducibility.…”

“…The most widely used in vitro models of the PC are based on a single protein [72,73], usually bovine serum albumin, or plasma in different variations and/or concentrations [4,71].…”

The effect of the protein corona on the interaction between nanoparticles and lipid bilayers

“…22 Since the PC affects the properties of the NP surface, it has significant impact on the interaction between membrane and NPs. [23][24][25][26][27] Many studies are described in the literature where NP cellular uptake is related to the presence of a PC on the NPs in biological fluids, [28][29][30] in particular it has been shown that a key factor in NPs cellular uptake is their adhesion to the cell membrane that significantly affects their final uptake rates. 31 NP-cell membrane interaction is a complex dynamic process in which the environmental proteins are known to play an important multifaceted role: not only they adsorb onto the NP surface forming the PC, but also they compete with the NP in the interaction with the cell membrane.…”

Particle Targeting in Complex Biological Media

“…Recruiting Method Performance Albumin BSA Non-specific adsorption Improved stability of NPs [161] Specific adsorption Prevented NPs from being filtered by the reticuloendothelial system in vivo [162] HSA Non-specific adsorption Enhanced the accessibility of targeting ligands and targeting results of functionalized polymeric particles; improved stability of NPs [87,94] Apolipo-protein ApoE Specific adsorption Helped particles to cross the BBB and to deliver therapeutics to the brain [104a,163] ApoJ (clusterin) Specific adsorption Created stealth surface of PEG-or PEEP-coated NPs and reduced non-specific cellular uptake of these NPs [59,123b] ApoA4 Specific adsorption Decreased cellular uptake of NPs [93] ApoC3 RBP Specific adsorption Directed NPs to hepatic stellate cells [162] Vitronectin Specific adsorption Promoted efficient uptake of particles in cancer cells expressing high levels of vitronectin α ν β 3 integrin receptor [164] Aβ 1-42 peptide Specific adsorption Captured toxic forms of Aβ 1-42 peptides and improved Alzheimer's disease condition [165] a)…”

“…[123b] Ritz et al reported that ApoA4 or ApoC3 precoated on particle surfaces significantly decreased the cellular uptake of particles, indicating the possibility of using these proteins as an endogenous alternative to PEG to mask non-specific bio-nano interactions. [93] By specifically recruiting or pre-adsorbing these endogenous proteins for surface functionalization, particles can display improved As researchers have become increasingly aware of the influence of adsorbed proteins from biological milieus, the recruitment of proteins to improve specific targeting of particles has been investigated. Recently, Santi et al showed that a rationally designed and in silico predicted peptide sequence tethered to Au NPs can specifically recruit and bind native Tf in a controlled orientation, while minimizing nonspecific protein adsorption from the biological environment.…”