Protein-coated Gold Nanoparticles Research Articles

Protein-Coated Aryl Modified Gold Nanoparticles for Cellular Uptake Study by Osteosarcoma Cancer Cells.

Gold nanoparticles coated with proteins have shown extraordinary biocompatibility which advanced to several nanomedicine engineering applications. We synthesized protein-coated gold nanoparticles using green and chemical reduction routes for cellular uptake study. In the current work, we coated gold-aryl nanoparticles of the type AuNPs-C6H4-4-COOH with bovine serum albumin (BSA), collagen, zein, and lysozyme proteins. Both routes were carried out without phase-transfer catalysts or extraneous stabilizing agents. High crystallinity of the AuNPs synthesized by the green route can be seen in transmission electron microscopy images. Osteosarcoma cancer cells are malignant bone tumors with abnormal cellular functions. Studies using MG-63 cells will provide mechanistic suggestions on the details of the amplification in tumors. We studied the cellular uptake of the bioconjugates by MG-63 osteosarcoma cells using laser confocal fluorescence microscopy (LCFM) and flow cytometry. In the LCFM study, BSA-AuNPs were uptaken most efficiently of all protein-coated gold nanoparticles synthesized by the green route. Lysozyme-AuNPs synthesized by the chemical reduction method were mostly efficiently internalized by MG-63 cells among all AuNPs. Zein- and lysozyme-coated AuNPs, though of relatively small size, prepared by the green method were not efficiently uptaken by MG-63. The two nanoparticles are negatively charged, and zein is also a hydrophobic coat. The difference in hydrophobicity and charge might have affected the internalization. All of those coated nanoparticles that were efficiently uptaken can potentially be used as diagnostic and therapeutic agents for osteosarcoma.

Synthesis of Metal@Protein@Polymer Nanoparticles with Distinct Interfacial and Phase Transfer Behavior

In this study, we present a novel and facile method for the synthesis of multiresponsive plasmonic nanoparticles with an interesting interfacial behavior. We used thiol-initiated photopolymerization technique to graft poly(N-isopropylacrylamide) onto the surface of protein-coated gold nanoparticles. The combination of the protein bovine serum albumin with the thermoresponsive polymer leads to smart hybrid nanoparticles, which show a stimuli-responsive behavior of their aggregation and a precisely controllable phase transfer behavior. Three interconnected stimuli, namely, temperature, ionic strength, and pH, were identified as property tuning switches. The aggregation was completely reversible and was quantified by determining Smoluchowski’s instability ratios with time-resolved dynamic light scattering. The tunable hydrophobicity via the three stimuli was used to study interfacial activity and phase transfer behavior of the nanoparticles at an octanol/water interface. Depending on the type of coating (i.e...

Protein coated gold nanoparticles as template for the directed synthesis of highly fluorescent gold nanoclusters

Bovine serum albumin (BSA) modified gold nanoparticles (AuNPs) was selected as template for the synthesis of AuNPs@gold nanoclusters (AuNCs) core/shell nanoparticles, in which BSA not only acted as dual functions agent for both anchoring and reducing Au3+ ions, but also was employed as a bridge between the AuNPs and AuNCs. Optical properties of AuNPs@AuNCs core/shell nanoparticles were studied using UV–visible and fluorescence spectroscopy. The prepared AuNPs@AuNCs core/shell nanoparticles exhibited sphere size uniformity with improved monodispersity, excellent fluorescence and fluorescent stability. Compared with AuNCs, AuNPs@AuNCs core/shell nanoparticles possessed large size and strong fluorescence intensity due to the effect of AuNPs as core. Moreover, the mechanism of the AuNPs induced fluorescence changes of the core/shell nanoparticles was first explored.

Taylor dispersion analysis, resonant mass measurement and bioactivity of pepsin-coated gold nanoparticles

Immobilized enzyme reactors based on nanoparticulate carriers are becoming increasingly popular. A toolbox of methods is usually utilized for their characterization in order to be capable of assessing their suitability for the intended purpose. In this work, as a model system pepsin was conjugated to gold nanoparticles (GNPs) by a straightforward adsorptive immobilization process. The success of the immobilization procedure was monitored by Vis spectroscopy via shifts of the localized surface plasmon resonance (LSPR) band, size characterization by dynamic light scattering (DLS), and ζ–potential determinations by electrophoretic light scattering. DLS revealed a significantly different hydrodynamic diameter for unmodified GNPs and protein-coated GNPs. However, the hydrodynamic diameters of pepsin-coated GNPs obtained with various concentrations of pepsin in the coating solution were not significantly different. In contrast, Taylor dispersion analysis allowed measuring the slight differences in the hydrodynamic radius. It provided also information on the viscosity of GNP suspensions and diffusion coefficients for the various pepsin@GNP preparations. For the determination of the pepsin surface coverage on the GNPs results from indirect protein quantitation of non-immobilized pepsin by Lowry assay were compared to direct measurement of immobilized pepsin by resonant mass measurements. Reasonable agreement was found. Accurate information on enzyme coverage is of utmost importance for a representative comparison of the turnover numbers and catalytic efficiencies of nanoparticulate immobilized enzyme reactors, which is shown by the adsorptively immobilized pepsin@GNP as model system.

Synergistic effect of reductase and keratinase for facile synthesis of protein-coated gold nanoparticles.

We have synthesized gold nanoparticles (GNPs) using chicken feathers (poultry waste) and Bacillus subtilis RSE163. Disulfide reductase and keratinase produced by Bacillus subtilis during the degradation of chicken feather has been used to reduce Au(3+) from HAuCl4 precursor to produce gold nanoparticles. The synthesized biogenic GNPs were characterized by UV-visible spectroscopy, transmission electron microscopy (TEM), and zeta potential measurements. Fourier transform infrared (FTIR) spectroscopy indicated the presence of protein capping on synthesized GNPs, imparting multifunctionality to the GNP surface. Furthermore, the nontoxic nature of biogenic GNPs was insured by interaction with Escherichia coli (ATCC11103), where TEM images and enhancement of growth rate of E. coli in log phase signified their nontoxic nature. The results indicate that the synthesis of biocompatible GNPs using poultry waste may find potential applications in drug delivery and sensing.

Strongly coupled plasmonic modes on macroscopic areas via template-assisted colloidal self-assembly.

We present ensembles of surface-ordered nanoparticle arrangements, which are formed by template-assisted self-assembly of monodisperse, protein-coated gold nanoparticles in wrinkle templates. Centimeter-squared areas of highly regular, linear assemblies with tunable line width are fabricated and their extinction cross sections can be characterized by conventional UV/vis/NIR spectroscopy. Modeling based on electrodynamic simulations shows a clear signature of strong plasmonic coupling with an interparticle spacing of 1–2 nm. We find evidence for well-defined plasmonic modes of quasi-infinite chains, such as resonance splitting and multiple radiant modes. Beyond elementary simulations on the individual chain level, we introduce an advanced model, which considers the chain length distribution as well as disorder. The step toward macroscopic sample areas not only opens perspectives for a range of applications in sensing, plasmonic light harvesting, surface enhanced spectroscopy, and information technology but also eases the investigation of hybridization and metamaterial effects fundamentally.

Surface plasmon resonances of protein-conjugated gold nanoparticles on graphitic substrates

We present theoretical calculations for the absorption properties of protein-coated gold nanoparticles on graphene and graphite substrates. As the substrate is far away from nanoparticles, numerical results show that the number of protein bovine serum molecules aggregating on gold surfaces can be quantitatively determined for gold nanoparticles with arbitrary size by means of the Mie theory and the absorption spectra. The presence of a graphene substrate near the protein-conjugated gold nanoparticles results in a red shift of the surface plasmon resonances of the nanoparticles. This effect can be modulated upon changing the graphene chemical potential. Our findings show that the graphene and graphite affect the absorption spectra in a similar way.

Physicochemical Properties of Protein‐Coated Gold Nanoparticles in Biological Fluids and Cells before and after Proteolytic Digestion

What's going on in there?! Little is known about the fate of nanoparticles (NPs) after their internalization by cells and organisms. Protein-coated gold NPs were used to study the physicochemical properties of NPs in extra- and intracellular fluids. These potential vehicles for enzymatic drug delivery were highly stable at pH 7.4 in the presence of salts and free proteins, but agglomerated reversibly under acidic conditions (see picture). As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Ultrafast Structural Dynamics of the Photocleavage of Protein Hybrid Nanoparticles

Protein-coated gold nanoparticles in suspension are excited by intense laser pulses to mimic the light-induced effect on biomolecules that occur in photothermal laser therapy with nanoparticles as photosensitizer. Ultrafast X-ray scattering employed to access the nanoscale structural modifications of the protein-nanoparticle hybrid reveals that the protein shell is expelled as a whole without denaturation at a laser fluence that coincides with the bubble formation threshold. In this ultrafast heating mediated by the nanoparticles, time-resolved scattering data show that proteins are not denatured in terms of secondary structure even at much higher temperatures than the static thermal denaturation temperature, probably because time is too short for the proteins to unfold and the temperature stimulus has vanished before this motion sets in. Consequently the laser pulse length has a strong influence on whether the end result is the ligand detachment (for example drug delivery) or biomaterial degradation.

The effect of cholesterol on protein-coated gold nanoparticle binding to liquid crystal-supported models of cell membranes

We report an easily visualized liquid crystal (LC)-based system to study the biophysical interactions between protein-coated gold nanoparticles (AuNPs) and LC-supported cell membrane model. The model consists of mixed phospholipid/cholesterol monolayer self-assembled at aqueous–LC interface. Protein-coated AuNPs were found to disrupt the mixed phospholipid/cholesterol monolayer. As a result, orientational transitions of LCs were triggered and optical responses of LCs from dark to bright were observed. The mixed monolayers with higher cholesterol contents were found to be more susceptible to the disruption by protein-coated AuNPs, and hydrophobic interaction played a major role in the monolayer disruption. We also found that the time for non-specific binding of fibrinogen-coated AuNPs to the mixed phospholipid/cholesterol monolayer was similar to that of specific binding of neutravidin-coated AuNPs to the mixed phospholipid/biotin-capped phospholipid monolayer. Results obtained from this study may offer new understanding in the potential nanotoxicity pathway, where the biophysical interaction between nanomaterials and cell membrane is an important step.

Imaging the disruption of phospholipid monolayer by protein-coated nanoparticles using ordering transitions of liquid crystals

We report an easily visualized liquid crystal (LC)-based system to study the molecular interactions between protein-coated gold nanoparticles (AuNPs) and supported phospholipid monolayer self-assembled at the aqueous-LC interface. Protein-coated AuNPs were found to disrupt the phospholipid monolayer and resulted in the orientational transitions of LCs that support the phospholipid layer. The disruption of the phospholipid monolayer depends on the type of protein (albumin, neutravidin, and fibrinogen) adsorbing onto nanoparticles. Furthermore, our results suggest that hydrophobic interaction plays a major role in the disruption of the phospholipid layer by protein-coated AuNPs. Results obtained from this study may offer new understanding in the potential cytotoxicity of nanomaterials, where the interaction between nanoparticles and cell membrane is an important step.

Elucidating the Mechanism of Cellular Uptake and Removal of Protein-Coated Gold Nanoparticles of Different Sizes and Shapes

We investigated the mechanism by which transferrin-coated gold nanoparticles (Au NP) of different sizes and shapes entered mammalian cells. We determined that transferrin-coated Au NP entered the cells via clathrin-mediated endocytosis pathway. The NPs exocytosed out of the cells in a linear relationship to size. This was different than the relationship between uptake and size. Furthermore, we developed a mathematical equation to predict the relationship of size versus exocytosis for different cell lines. These studies will provide guidelines for developing NPs for imaging and drug delivery applications, which will require "controlling" NP accumulation rate. These studies will also have implications in determining nanotoxicity.

Gold Nanoparticles as a Colorimetric Sensor for Protein Conformational Changes

Spherical gold nanoparticles and flat gold films are prepared in which yeast iso-1-cytochrome c (Cyt c) is covalently bound to the gold surface by a thiol group in the cystein 102 residue. Upon exposure to solutions of different pH, bound Cyt c unfolds at low pH and refolds at high pH. This conformational change causes measurable shifts in the color of the coated nanoparticle solutions detected by UV-VIS absorption spectroscopy and in the refractive index (RI) of the flat gold films detected by surface plasmon resonance (SPR) spectroscopy. Both experiments demonstrate the same trend with pH, suggesting the use of protein-covered gold nanoparticles as a simple colorimetric sensor for conformational change.