Nanoparticle Corona Research Articles

Dual Fractions Proteomic Analysis of Silica Nanoparticle Interactions with Protein Extracts

Dual-fraction proteomics reveals a novel class of proteins impacted by nanoparticle exposure. Background: Nanoparticles (NPs) interact with cellular proteomes, altering biological processes. Understanding these interactions requires comprehensive analyses beyond solely characterizing the NP corona. Methods: We utilized a dual-fraction mass spectrometry (MS) approach to analyze both NP-bound and unbound proteins in Saccharomyces cerevisiae sp. protein extracts exposed to silica nanoparticles (SiNPs). We identified unique protein signatures for each fraction and quantified protein abundance changes using spectral counts. Results: Strong correlations were observed between protein profiles in each fraction and non-exposed controls, while minimal correlation existed between the fractions themselves. Linear models demonstrated equal contributions from both fractions in predicting control sample abundance. Combining both fractions revealed a larger proteomic response to SiNP exposure compared to single-fraction analysis. We identified 302/56 proteins bound/unbound to SiNPs and an additional 196 “impacted” proteins demonstrably affected by SiNPs. Conclusion: This dual-fraction MS approach provides a more comprehensive understanding of nanoparticle interactions with cellular proteomes. It reveals a novel class of “impacted” proteins, potentially undergoing conformational changes or aggregation due to NP exposure. Further research is needed to elucidate their biological functions and the mechanisms underlying their impact.

Protein Corona of Nanoparticles: Isolation and Analysis

Protein Corona of Nanoparticles: Isolation and Analysis

Mass Spectrometry-Based Top-Down Proteomics in Nanomedicine: Proteoform-Specific Measurement of Protein Corona.

Conventional mass spectrometry (MS)-based bottom-up proteomics (BUP) analysis of the protein corona [i.e., an evolving layer of biomolecules, mostly proteins, formed on the surface of nanoparticles (NPs) during their interactions with biomolecular fluids] enabled the nanomedicine community to partly identify the biological identity of NPs. Such an approach, however, fails to pinpoint the specific proteoforms─distinct molecular variants of proteins in the protein corona. The proteoform-level information could potentially advance the prediction of the biological fate and pharmacokinetics of nanomedicines. Recognizing this limitation, this study pioneers a robust and reproducible MS-based top-down proteomics (TDP) technique for characterizing proteoforms in the protein corona. Our TDP approach has successfully identified about 900 proteoforms in the protein corona of polystyrene NPs, ranging from 2 to 70 kDa, revealing proteoforms of 48 protein biomarkers with combinations of post-translational modifications, signal peptide cleavages, and/or truncations─details that BUP could not fully discern. This advancement in MS-based TDP offers a more advanced approach to characterize NP protein coronas, deepening our understanding of NPs' biological identities. We, therefore, propose using both TDP and BUP strategies to obtain more comprehensive information about the protein corona, which, in turn, can further enhance the diagnostic and therapeutic efficacy of nanomedicine technologies.

Nanoparticle-assisted tubulin assembly is environment dependent

Nanomaterials acquire a biomolecular corona upon introduction to biological media, leading to biological transformations such as changes in protein function, unmasking of epitopes, and protein fibrilization. Ex vivo studies to investigate the effect of nanoparticles on protein-protein interactions are typically performed in buffer and are rarely measured quantitatively in live cells. Here, we measure the differential effect of silica nanoparticles on protein association in vitro vs. in mammalian cells. BtubA and BtubB are a pair of bacterial tubulin proteins identified in Prosthecobacter strains that self-assemble like eukaryotic tubulin, first into dimers and then into microtubules in vitro or in vivo. Förster resonance energy transfer labeling of each of the Btub monomers with a donor (mEGFP) and acceptor (mRuby3) fluorescent protein provides a quantitative tool to measure their binding interactions in the presence of unfunctionalized silica nanoparticles in buffer and in cells using fluorescence spectroscopy and microscopy. We show that silica nanoparticles enhance BtubAB dimerization in buffer due to protein corona formation. However, these nanoparticles have little effect on bacterial tubulin self-assembly in the complex mammalian cellular environment. Thus, the effect of nanomaterials on protein-protein interactions may not be readily translated from the test tube to the cell in the absence of particle surface functionalization that can enable targeted protein-nanoparticle interactions to withstand competitive binding in the nanoparticle corona from other biomolecules.

A magnetic separation method for isolating and characterizing the biomolecular corona of lipid nanoparticles

Lipid nanoparticle (LNP) formulations are a proven method for the delivery of nucleic acids for gene therapy as exemplified by the worldwide rollout of LNP-based RNAi therapeutics and mRNA vaccines. However, targeting specific tissues or cells is still a major challenge. After LNP administration, LNPs interact with biological fluids (i.e., blood), components of which adsorb onto the LNP surface forming a layer of biomolecules termed the "biomolecular corona (BMC)" which affects LNP stability, biodistribution, and tissue tropism. The mechanisms by which the BMC influences tissue- and cell-specific targeting remains largely unknown, due to the technical challenges in isolating LNPs and their corona from complex biological media. In this study, we present a new technique that utilizes magnetic LNPs to isolate LNP-corona complexes from unbound proteins present in human serum. First, we developed a magnetic LNP formulation, containing >40 superparamagnetic iron oxide nanoparticles (IONPs)/LNP, the resulting LNPs containing iron oxide nanoparticles (IOLNPs) displayed a similar particle size and morphology as LNPs loaded with nucleic acids. We further demonstrated the isolation of the IOLNPs and their corresponding BMC from unbound proteins using a magnetic separation (MS) system. The BMC profile of LNP from the MS system was compared to size exclusion column chromatography and further analyzed via mass spectrometry, revealing differences in protein abundances. This new approach enabled a mild and versatile isolation of LNPs and its corona, while maintaining its structural integrity. The identification of the BMC associated with an intact LNP provides further insight into LNP interactions with biological fluids.

Protein Binding Leads to Reduced Stability and Solvated Disorder in the Polystyrene Nanoparticle Corona.

Understanding the conformation of proteins in the nanoparticle corona has important implications in how organisms respond to nanoparticle-based drugs. These proteins coat the nanoparticle surface, and their properties will influence the nanoparticle's interaction with cell targets and the immune system. While some coronas are thought to be disordered, two key unanswered questions are the degree of disorder and solvent accessibility. Here, a model is developed for protein corona disorder in polystyrene nanoparticles of varying size. For two different proteins, it is found that binding affinity decreases as nanoparticle size increases. The stoichiometry of binding, along with changes in the hydrodynamic size, supports a highly solvated, disordered protein corona anchored at a small number of attachment sites. The scaling of the stoichiometry versus nanoparticle size is consistent with disordered polymer dimensions. Moreover, it is found that proteins are destabilized less in the presence of larger nanoparticles, and hydrophobic exposure decreases at lower curvatures. The observations hold for proteins on flat polystyrene surfaces, which have the lowest hydrophobic exposure. The model provides an explanation for previous observations of increased amyloid fibrillation rates in the presence of larger nanoparticles, and it may rationalize how cell receptors can recognize protein disorder in therapeutic nanoparticles.

Exploring Residue-Level Interactions between the Biofilm-Driving R2ab Protein and Polystyrene Nanoparticles.

In biological systems, proteins can bind to nanoparticles to form a "corona" of adsorbed molecules. The nanoparticle corona is of significant interest because it impacts an organism's response to a nanomaterial. Understanding the corona requires knowledge of protein structure, orientation, and dynamics at the surface. A residue-level mapping of protein behavior on nanoparticle surfaces is needed, but this mapping is difficult to obtain with traditional approaches. Here, we have investigated the interaction between R2ab and polystyrene nanoparticles (PSNPs) at the level of individual residues. R2ab is a bacterial surface protein from Staphylococcus epidermidis and is known to interact strongly with polystyrene, leading to biofilm formation. We have used mass spectrometry after lysine methylation and hydrogen-deuterium exchange (HDX) NMR spectroscopy to understand how the R2ab protein interacts with PSNPs of different sizes. Lysine methylation experiments reveal subtle but statistically significant changes in methylation patterns in the presence of PSNPs, indicating altered protein surface accessibility. HDX rates become slower overall in the presence of PSNPs. However, some regions of the R2ab protein exhibit faster than average exchange rates in the presence of PSNPs, while others are slower than the average behavior, suggesting conformational changes upon binding. HDX rates and methylation ratios support a recently proposed "adsorbotope" model for PSNPs, wherein adsorbed proteins consist of unfolded anchor points interspersed with partially structured regions. Our data also highlight the challenges of characterizing complex protein-nanoparticle interactions using these techniques, such as fast exchange rates. While providing insights into how R2ab adsorbs onto PSNP surfaces, this research emphasizes the need for advanced methods to comprehend residue-level interactions in the nanoparticle corona.

Looking back, moving forward: protein corona of lipid nanoparticles.

Lipid nanoparticles (LNPs) are commonly employed for drug delivery owing to their considerable drug-loading capacity, low toxicity, and excellent biocompatibility. Nevertheless, the formation of protein corona (PC) on their surfaces significantly influences the drug's in vivo fate (such as absorption, distribution, metabolism, and elimination) upon administration. PC denotes the phenomenon wherein one or multiple strata of proteins adhere to the external interface of nanoparticles (NPs) or microparticles within the biological milieu, encompassing ex vivo fluids (e.g., serum-containing culture media) and in vivo fluids (such as blood and tissue fluids). Hence, it is essential to claim the PC formation behaviors and mechanisms on the surface of LNPs. This overview provided a comprehensive examination of crucial aspects related to such issues, encompassing time evolution, controllability, and their subsequent impacts on LNPs. Classical studies of PC generation on the surface of LNPs were additionally integrated, and its decisive role in shaping the in vivo fate of LNPs was explored. The mechanisms underlying PC formation, including the adsorption theory and alteration theory, were introduced to delve into the formation process. Subsequently, the existing experimental outcomes were synthesized to offer insights into the research and application facets of PC, and it was concluded that the manipulation of PC held substantial promise in the realm of targeted delivery.

Nanoparticle Isolation from Biological Media for Protein Corona Analysis: The Impact of Incubation and Recovery Protocols on Nanoparticle Properties

Nanoparticles are increasingly implemented in biomedical applications, including the diagnosis and treatment of disease. When exposed to complex biological media, nanoparticles spontaneously interact with their surrounding environment, leading to the surface-adsorption of small and bio- macromolecules- termed the “corona”. Corona composition is governed by nanoparticle properties and incubation parameters. While the focus of most studies is on the protein signature of the nanoparticle corona, the impact of experimental protocols on nanoparticle size in the presence of complex biological media, and the impact of nanoparticle recovery from biological media has not yet been reported. Here using a non-degradable robust model, we show how centrifugation-resuspension protocols used for the isolation of nanoparticles from incubation media, incubation duration and shear flow conditions alter nanoparticle parameters including particle size, zeta potential and total protein content.Our results show significant changes in nanoparticle size following exposure to media containing protein under different flow conditions, which also altered the composition of surface-adsorbed proteins profiled by SDS-PAGE. Our in situ analysis of nanoparticle size in media containing protein using particle tracking analysis highlights that centrifugation-resuspension is disruptive to agglomerates that are spontaneously formed in protein containing media, highlighting the need for in situ analytical methods that do not alter the intermediates formed following nanoparticle exposure to biological media.Nanomedicines are mostly intended for parenteral administration, and our findings show that parameters such as shear flow can significantly alter nanoparticle physicochemical parameters. Overall, we show that the centrifugation-resuspension isolation of nanoparticles from media significantly alters particle parameters in addition to the overall protein composition of surface-adsorbed proteins. We recommend that nanoparticle characterization pipelines studying bio-nano interactions during early nanomedicine development consider biologically-relevant shear flow conditions and media composition that can significantly alter particle physical parameters and subsequent conclusions from these studies.

Visualizing the Birth and Monitoring the Life of a Bimetallic Methanation Catalyst.

Well-defined bimetallic heterogeneous catalysts are not only difficult to synthesize in a controlled manner, but their elemental distributions are also notoriously challenging to define. Knowledge of these distributions is required for both the as-synthesized catalyst and its activated form under reaction conditions, where various types of reconstruction can occur. Success in this endeavor requires observation of the active catalyst via in situ analytical methods. As a step toward this goal, we present a composite material composed of bimetallic nickel-ruthenium nanoparticles supported on a protonated zeolite (Ni-Ru/HZSM-5) and probe its evolution and function as a photoactive carbon dioxide methanation catalyst using in situ X-ray absorption spectroscopy (XAS). The working Ni-Ru/HZSM-5, as a selective and durable photothermal CO2 methanation catalyst, comprises a corona of Ru nanoparticles decorating a Ni nanoparticle core. The specific Ni-Ru interactions in the bimetallic particles were confirmed by in situ XAS, which reveals significant electron transfer from Ni to Ru. The light-harvesting Ni nanoparticle core and electron-accepting Ru nanoparticle corona serve as the CO2 and H2 dissociation centers, respectively. These Ni and Ru nanoparticles also promote synergistic photothermal and hydrogen atom transfer effects. Collectively, these effects enable an associative CO2 methanation reaction pathway while hindering coking and fostering high selectivity toward methane.

Ectoenzymes as promising cell identification structures for the high avidity targeting of polymeric nanoparticles

Pharmacotherapy is often limited by undesired side effects while insufficient drug reaches the site of action. Active-targeted nanotherapy should provide a solution for this problem, by using ligands in the nanoparticle corona for the identification of receptors on the target-cell surface. However, since receptor binding is directly associated with pharmacological responses, today's targeting concepts must be critically evaluated. We hypothesized that addressing ectoenzymes would help to overcome this problem, but it was not clear if particles would show sufficiently high avidity to provide us with a viable alternative to classical ligand-receptor concepts. We scrutinized this aspect by immobilizing the highly selective angiotensin-converting enzyme 2 (ACE2) inhibitor MLN-4760 in the corona of block-copolymer nanoparticles and investigated enzyme binding via microscale thermophoresis and flow cytometry. Excellent avidities with Kd values as low as 243 pM for soluble ACE2 and 306 pM for ACE2-positive cells were obtained. In addition, the inhibitory activity had an IC50 value of 2.88 nM. Reliable target cell identification could be proven in coculture experiments. High avidity is the basis for minimizing material loss to off-target sites and paves the way for a paradigm shift in nanoparticle targeting which does not trigger unintended side effects following target cell identification.

Biophysical study of DC electric field induced stable formation of albumin-gold nanoparticles corona and curcumin binding

Targeted drug delivery (TDD) is a method of delivering optimum concentrations of pharmaceutical substances in the tissue to achieve the desired therapeutic effect. Hence, TDD systems are considered as an emerging strategy to deliver the drug at the specific site of the tissues/cells. The nanoparticle-protein corona as a drug delivery vehicle has demonstrated immense benefits including potential theragnostic, improved pharmacodynamics and targeted drug delivery. In the present investigation, efforts have been to establish stable and functionalized Bovine serum albumin–gold nanoparticle (BSA-GNP) corona (conjugates) using a Direct Current (DC) electric field. With the application of DC electric field (DEF) across the BSA-GNP solution, the formation of BSA-GNP corona/conjugate takes place which was characterized using various biophysical techniques such a Dynamic Light Scattering (DLS), UV Visible spectroscopy, Fluorescence spectroscopy, electrophoresis, etc. Furthermore, the DEF engineered BSA-GNP corona was loaded/interacted with curcumin (CUR). The size of the BSA-GNP corona was increased with increasing DC voltage (5–30 V) at constant concentration of BSA. The strong and stable binding of curcumin with BSA-GNP corona was revealed by the techniques used in the investigation; however, binding affinity of CUR was decreased for 30 V DEF exposed BSA-GNP conjugate. The biocompatible experimental data confirms the nontoxic nature of BSA-GNP corona. This investigation adds a new and novel physical method for the preparation of protein-nanoparticle corona for various applications including drug delivery.

Identification and characterization of soft protein corona absorbed on iron oxide nanoparticles

Nanomedicines have recently garnered significant attention due to their great potential to improve the efficiency of disease diagnosis and treatment. Nonetheless, a wide gap still exists between innovative research and clinical application because of limited understanding of the protein corona formed on the nanocarrier's surface. When NPs are introduced to living systems, the protein corona rather than the originally expected surface of the nanoparticle is initially recognized, further affecting immune clearance and tumor-specific delivery. This study found that after incubation with human plasma, the protein corona formed on the iron oxide nanoparticle's (IONP) surface significantly increased the uptake of IONPs in the leukemic cell line MOLM13. The formation and thicknesses of the protein coronas were measured by dynamic light scattering, nanoparticle tracking analysis, and flow cytometry, and the hard corona was measured by transmission electron microscopy. The size distribution of IONPs before and after incubation with plasma was measured, and the change in particle size was calculated to derive the thickness of the soft corona (7.58±3.94 nm) by subtracting the thickness of the hard corona from that of the full protein corona. The results can provide an approximation of the soft corona area after plasma incubation and contribute to the understanding of the features of the protein corona of nanoparticles with different physical and chemical properties.

Impact of Nanoparticle Physicochemical Properties on Protein Corona and Macrophage Polarization.

Macrophages, the major component of the mononuclear phagocyte system, uptake and clear systemically administered nanoparticles (NPs). Therefore, leveraging macrophages as a druggable target may be advantageous to enhance NP-mediated drug delivery. Despite many studies focused on NP-cell interactions, NP-mediated macrophage polarization mechanisms are still poorly understood. This work aimed to explore the effect of NP physicochemical parameters (size and charge) on macrophage polarization. Upon exposure to biological fluids, proteins rapidly adsorb to NPs and form protein coronas. To this end, we hypothesized that NP protein coronas govern NP-macrophage interactions, uptake, and subsequent macrophage polarization. To test this hypothesis, model polystyrene NPs with various charges and sizes, as well as NPs relevant to drug delivery, were utilized. Data suggest that cationic NPs potentiate both M1 and M2 macrophage markers, while anionic NPs promote M1-to-M2 polarization. Additionally, anionic polystyrene nanoparticles (APNs) of 50 nm exhibit the greatest influence on M2 polarization. Proteomics was pursued to further understand the effect of NPs physicochemical parameters on protein corona, which revealed unique protein patterns based on NP charge and size. Several proteins impacting M1 and M2 macrophage polarization were identified within cationic polystyrene nanoparticles (CPNs) corona, while APNs corona included fewer M1 but more M2-promoting proteins. Nevertheless, size impacts protein corona abundance but not identities. Altogether, protein corona identities varied based on NP surface charge and correlated to dramatic differences in macrophage polarization. In contrast, NP size differentially impacts macrophage polarization, which is dominated by NP uptake level rather than protein corona. In this work, specific corona proteins were identified as a function of NP physicochemical properties. These proteins are correlated to specific macrophage polarization programs and may provide design principles for developing macrophage-mediated NP drug delivery systems.

Polyelectrolyte nanoparticles based on poly[N-(2-hydroxypropyl)methacrylamide-block-poly(N-(3-aminopropyl)methacrylamide] copolymers for delivery of heparin-binding proteins

In addition to the delivery of therapeutic nucleic acids and low-molecular weight drugs, polyelectrolyte nanoparticles (NPs) have recently been studied as carriers for protein delivery. However, the stability of NPs during isolation steps, which ensures their easy redispersion, needs to be solved empirically for individual systems using cryoprotectants and stabilizers. To avoid the use of additives, we studied the formation of polyelectrolyte NPs consisting of a newly synthesized polycationic diblock copolymer based on poly(N-(2-hydroxypropyl)methacrylamide)-block-poly(N-(3-aminopropyl)methacrylamide) (p(HPMA-b-APMA) and heparin (Hep). The p(APMA) blocks electrostatically complexed with Hep, and the p(HPMA) blocks formed a neutral corona of NPs, limiting NP aggregation. Self-assembly was monitored through the changes in size and zeta potential of the formed NPs, which depended on the copolymer composition and the concentration of polyelectrolyte solutions. The interactions between the NP components were analysed by FTIR spectroscopy, and XPS analysis indicated the presence of p(HPMA) blocks on the surface of the NPs. The encapsulation of the model chemokine CXCL12 and basic fibroblast growth factor (FGF-2) was driven by their specific bioaffinity for Hep, resulting in a high 90 % entrapment efficiency and increased protein stability. CXCL12 was released over 48 h, while FGF-2 exhibited a sustained release of up to 38 % over four weeks. In addition, the released CXCL12 effectively stimulated the migration of macrophages and T-lymphocyte cells, indicating the preserved protein bioactivity. Considering the proven noncytotoxic performance of the NPs towards fibroblasts and mesenchymal stem cells, the polyelectrolyte NPs of p(HPMA-b-APMA) and Hep loaded with heparin-binding proteins can be considered as promising candidates for the controlled delivery of bioactive proteins in biomedical applications.

An artificialed protein corona coating the surface of magnetic nanoparicles:a simple and efficient method for label antibody

BackgroundProtein Corona (PC) of nanoparticles is a structure which composed of one or more layers of proteins adsorbed on the surface of nanomaterials, and the formation of PC is a universal process of spontaneous randomness. We take advantage of the formation principle of the PC, developed a simple and efficient method for label protein to nanoparticles. MethodsThe artificialed protein corona (APC) on the surface of nanoparticles was synthesized via the artificialed methods of desolvation aggregation and crosslinking with control. ResultsThe dosage of precipitator and the ratio of protein to magnetic nanoparticles (MNPs)(particle size: 3 nm) were optimized, and the core-shell nanoparticles with narrow particle size (particle size: 10 nm) distribution were obtained. The MNPs with APC were characterized by transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM). Additionally, a hemolysis test on prepared MNPs was conducted with APC. The presence of APC coating on the surface of MNPs showed an improving effect to reduce the cytotoxicity. Cellular toxicity of MNPs with APC was also investigated on HFF1 cell lines. And the cells survival in the presence of APC coated MNPs and display neither reduced metabolism nor cytostatic effect. The functional test of the MNPs with APC showed that proteins can be modified and labeled onto magnetic nanoparticles and retain their original activity. ConclusionsThis marking method is gentle and effective. And the properties of the APC propose MNPs as a promising candidate for multifunctional biomedical applications.

Emergence of microbial resistance against nanoparticles: Mechanisms and strategies.

Antimicrobial nanoparticles have gained the status of a new generation of drugs that can kill bacterial pathogens by multiple means; however, nanoparticle resistance acquired by some bacterial pathogens has evoked a cause of concern. Several reports suggested that bacteria can develop nanoparticles, specifically metal nanoparticle resistance, by mechanisms: nanoparticle transformation-induced oxidative stress, membrane alterations, reversible adaptive resistance, irreversible modifications to cell division, and a change in bacterial motility and resistance. Surface properties, concentration and aggregation of nanoparticles, biofilm forming and metal exclusion capacity, and R plasmid and flagellin synthesis by bacteria are crucial factors in the development of nanoparticle resistance in bacteria. Studies reported the resistance reversal by modifying the surface corona of nanoparticles or inhibiting flagellin production by bacterial pathogens. Furthermore, strict regulation regarding the use and disposal of nano-waste across the globe, the firm knowledge of microbe-nanoparticle interaction, and the regulated disposal of nanoparticles in soil and water is required to prevent microbes from developing nanoparticle resistance.

Encapsulation of phenylacetic acid in block copolymer nanoparticles during polymerization induced self‐assembly

AbstractPolymerization induced self‐assembly (PISA) is an in situ method for producing block copolymer nanoparticles. Performing PISA in the presence of a pharmaceutical drug causes the nanoparticles to encapsulate the drug. While this approach is straightforward, the effects of drug loading and block copolymer composition remain unclear. Here, we investigate encapsulation of the drug phenylacetic acid (PA) in poly(glycerol monomethacrylate)‐block‐poly(2‐hydroxypropyl methacrylate) (PGMA‐PHPMA) nanoparticles during PISA. Nanoparticle morphology is characterized by electron microscopy and light scattering, while encapsulation efficiency () is quantified using nuclear magnetic resonance diffusometry. Increasing the PA loading shifts the nanoparticle morphology from spherical micelles cylindrical micelles vesicles. At a 32 mg/ml PA loading, maximizes at ~80%. Increasing the PHPMA degree of polymerization minimally impacts . The invariance of toward core block length suggests that PA binds to the nanoparticle corona, highlighting the importance of the hydrophilic block for drug encapsulation during PISA.

Interactions of Apolipoproteins with Lipid-Based Nanoparticles.

Serum proteins bind and form a dynamic protein corona around nanoparticles (NPs) that have been injected into the mammalian vasculature. Several fundamental studies have shown that apolipoproteins are prominent components of the NP corona. Since apolipoproteins control the distribution of lipoproteins, they may also control the distribution of NPs. Indeed, apolipoprotein affinity for NPs has been recently taken advantage of to deliver CRISPR reagents encapsulated in NPs to cells that express particular lipoprotein receptors. In this scenario, an apolipoprotein binds an NP and the resulting apolipoprotein-NP complex binds a cell that expresses the (apo)lipoprotein receptor. But the NP will be diverted from the target cell if it does not express the (apo)lipoprotein receptor. This may hamper NP treatment of diseases. Therefore, we must understand the kinetics of apolipoprotein-NP affinity and how apolipoprotein-NP interactions affect NP biodistribution. In this Perspective, we discuss the evolving topic of apolipoprotein-NP interactions, which is of great interest for all NP-based disease treatments. Many properties of apolipoprotein-NP complexes are yet to be determined and will have a significant impact on NP efficacy for many NP-based treatments in animal models and in the clinic.

Probing the Protein Corona of Nanoparticles in a Fluid Flow by Single-Particle Differenced Resonance Light Scattering Correlation Spectroscopy.

The protein corona of nanoparticles (NPs) plays a crucial role in determining NPs' biological fates. Here, a novel measurement strategy was proposed to in situ investigate the protein corona formed in the NPs with the home-built dual-wavelength laser-irradiated differenced resonance light scattering correlation spectroscopy (D-RLSCS) technique, combined with the modified generation method of the D-RLSCS curve. With the measurement strategy, the dissociation constants and the binding rates between proteins and gold nanoparticles (GNPs) were determined based on the binding-induced ratiometric diffusion change of NPs (the ratio of characteristic rotational diffusion time to translational one), using the formation of the protein corona of bovine serum albumin (BSA) or fibrinogen (FIB) on gold nanoparticles as a model. It was found that BSA shows a stronger binding constant and faster binding rate to gold nanospheres (GNSs) compared with those of FIB. Meanwhile, the dynamic behavior of the protein corona in a fluid flow mimicking biological vessels was further studied based on the combination of the D-RLSCS technique with a microfluidic channel. The measurement results indicated that some "loose" protein corona layers would strip off the surface of NPs within the microchannel due to the fluid sheath force. This method can provide the comprehensive information of a protein corona by averaging the diffusion behavior of many particles different from some conventional methods and overcome the shortcomings of conventional correlation spectroscopy methods.