Impact of Protein Corona in Nanoflare-Based Biomolecular Detection and Quantification

Abstract

Selective detection and precise quantification of biomolecules in intracellular settings play a pivotal role in the diagnostics and therapeutics of diseases, including various cancers and infectious epidemics. Because of this clinical relevance, nanoprobes with high sensitivity, wide tunability, and excellent biological stability have become of high demand. In particular, nanoflares based on gold nanoparticles have emerged as an attractive candidate for intracellular detection due to their efficient cellular uptake, enhanced binding affinity with complementary targets, and improved biological compatibility. However, nanoprobes, including these nanoflares, are known to be susceptible to the adsorption of proteins present in the biological environment, which leads to the formation of a so-called protein corona layer on their surface, leading to an altered targeting efficiency and cellular uptake. In this work, we leverage the nanoflares platform to demonstrate the effect of protein corona on biomolecular detection, quantification, as well as biological stability against enzymatic degradation. Nanoflares incubated in a biologically relevant concentration of serum albumin proteins (0.50 wt %) were shown to result in more than 20% signal reduction in target detection, with a decrease varying proportionally with the protein concentrations. In addition, similar signal reduction was observed for different serum proteins, and PEG backfilling was found to be ineffective in mitigating the negative impact induced by the corona formation. Furthermore, nuclease resistance in nanoflares was also severely compromised by the presence of the corona shell (∼2-fold increase in hydrolysis activity). This work demonstrates the consequences of an in situ formed protein corona layer on molecular detection/quantification and biological stability of nanoflares in the presence of nuclease enzymes, highlighting the importance of calibrating similar nanoprobes in proper biological media to improve the accuracy of molecular detection and quantification.

Rapid point‐of‐need detection of bacteria and their toxins in food using gold nanoparticles

“…Its thickness and structure depends on the medium composition but also on the NPs' physicochemical properties and time of exposure (García-Álvarez et al, 2018;Piella et al, 2017). Wang et al, have shown that a protein corona created by albumin adsorption on AuNPs decreased the detection signal of the LSPR sensor by more than 20% compared to the detection signal observed in a buffer solution containing no protein (H. Wang et al, 2019). Most research on protein corona formed around AuNPs has been performed using biological fluids or cell media with only a very few dealing with this point using food matrices.…”

Spherical Nucleic Acids with Tailored and Active Protein Coronae

“…However, many nanomaterials, when introduced to biological fluids, nonspecifically adsorb biomolecules, resulting in the formation of a protein corona around the structure . The protein corona alters the biological stability, − biodistribution, , and targeting efficiency − of a nanomaterial, sometimes diminishing its therapeutic potential. Though the surface charge, , size, ,, and shape of a nanomaterial can modulate the composition of the protein corona, its formation is largely unavoidable in biological environments.…”

Engineering the Interface between Inorganic Nanoparticles and Biological Systems through Ligand Design

“…Nanoparticles administered into the blood can interact nonspecifically with serum proteins and form a shell of protein, called the protein corona [39][40][41][42][43][44]. Protein corona formation is mainly mediated by coulombic and Van der Waals forces, hydrogen bonding, and hydrophobic interactions [45].…”