Entropy-Driven Reversible Agglomeration of Crown Ether Capped Gold Nanoparticles.

It is shown that plasmonic gold nanoparticles functionalised with a thiolated 18-crown-6 ligand shell agglomerate spontaneously from aqueous dispersion at elevated temperatures. This process takes place over a narrow temperature range, is accompanied by a colour change from red to purple-blue and is fully reversible. Moreover, the temperature at which it occurs can be adjusted by the degree of complexation of the crown ether moiety with appropriate cations. More complexation leads to higher transition temperatures. The process has been studied by UV/Vis spectroscopy, electron microscopy, dynamic light scattering and zeta potential measurements. A thermodynamic rationale is provided to suggest an entropy-driven endothermic agglomeration process based on attractive hydrophobic interactions of the complexed crowns that are competing against electrostatic repulsion of the charged ligand shells.

Sensitive Analysis of Protein Adsorption to Colloidal Gold by Differential Centrifugal Sedimentation.

It is demonstrated that the adsorption of bovine serum albumin (BSA) to aqueous gold colloids can be quantified with molecular resolution by differential centrifugal sedimentation (DCS). This method separates colloidal particles of comparable density by mass. When proteins adsorb to the nanoparticles, both their mass and their effective density change, which strongly affects the sedimentation time. A straightforward analysis allows quantification of the adsorbed layer. Most importantly, unlike many other methods, DCS can be used to detect chemisorbed proteins ("hard corona") as well as physisorbed proteins ("soft corona"). The results for BSA on gold colloid nanoparticles can be modeled in terms of Langmuir-type adsorption isotherms (Hill model). The effects of surface modification with small thiol-PEG ligands on protein adsorption are also demonstrated.

Design of artificial membrane transporters from gold nanoparticles with controllable hydrophobicity.

Gold nanoparticles with variable hydrophobicity have been prepared in three different size regimes following established methods. The control of hydrophobicity was achieved by complexation of the 18-crown-6-CH2-thiolate ligand shell with potassium ions. Potassium dependent phase transfer of these particles from dispersion in water to chloroform was demonstrated, and the equilibrium partitioning of the particles in water-chloroform liquid/liquid systems was quantified by optical spectroscopy. The gradual complexation of the ligand shell with potassium ions was further monitored by zeta potential measurements. Potassium dependent insertion of nanoparticles into the phospholipid bilayer membrane of vesicles in aqueous dispersion has been demonstrated by cryogenic transmission electron microscopy (cryo-TEM). Nanoparticle-dependent potassium ion transport across the vesicle membrane has been established by monitoring the membrane potential with fluorescence spectroscopy using a potential sensitive dye.

Electrocatalytic hydrogen redox chemistry on gold nanoparticles.

Electrocatalytic proton reduction leading to the formation of adsorbed molecular hydrogen on gold nanoparticles of 1-3 and 14-16 nm diameter stabilized by 1-mercapto-undecane-11-tetra(ethyleneglycol) has been demonstrated by cyclic voltammetry using a hanging mercury drop electrode. The nanoparticles were adsorbed to the electrode from aqueous dispersion and formed robust surface layers transferrable to fresh base electrolyte solutions. Unique electrocatalytic proton redox chemistry was observed that has no comparable counterpart in the electrochemistry of bulk gold electrodes. Depending on size, the nanoparticles have a discrete number of electrocatalytically active sites for the two-electron/two-proton reduction process. The adsorbed hydrogen formed is oxidized with the reverse potential sweep. These findings represent a new example of qualitative different behavior of nanoparticles in comparison with the corresponding bulk material.

Mercaptocarborane-capped gold nanoparticles: electron pools and ion traps with switchable hydrophilicity.

A simple single-phase method for the preparation of ca. 2 nm gold nanoparticles capped with mercaptocarborane ligands is introduced. The resultant monolayer protected clusters (MPCs) exhibit redox-dependent solubility and readily phase transfer between water and nonpolar solvents depending on the electronic and ionic charge stored in the metal core and in the ligand shell, respectively. The particles and their properties have been characterized by high angle annular dark field imaging in a scanning transmission electron microscope, elemental analysis, centrifugal particle sizing, UV-vis and FTIR spectroscopy, and thermogravimetric analysis and by (1)H, (11)B, and (7)Li NMR spectroscopy. Cellular uptake of the MPCs by HeLa cells has been studied by TEM, and the subsequent generation of reactive oxygen species inside the cells has been evaluated by confocal fluorescence microscopy. These MPCs qualitatively showed significant toxicity and the ability to penetrate into most cell compartments with a strong tendency of finally residing inside membranes. Applications in catalysis, electrocatalysis, and biomedicine are envisaged.

Inflicting controlled nonthermal damage to subcellular structures by laser-activated gold nanoparticles.

We show that low-intensity laser irradiation of cancer cells containing endosomal gold nanoparticles leads to endosome rupture and escape of the nanoparticles into the cytosol without affecting the cells' viability. The low light intensity of our experiments allows us to rule out photothermal effects as the underlying mechanism, and we present results that suggest photoinduced radicals as the photogenerated active species. This nonthermal mechanism may also be important in the context of cell death at higher laser intensities, which had been reported previously.

Ion shuttling between emulsion droplets by crown ether modified gold nanoparticles.

Selective unidirectional transport of barium ions between droplets in a water-in-chloroform emulsion is demonstrated. Gold nanoparticles (GNPs) modified with a thiolated crown ether act as barium ion complexing shuttles that carry the ions from one population of droplets (source) to another (target). This process is driven by a steep barium ion concentration gradient between source and target droplets. The concentration of barium ions in the target droplets is kept low at all times by the precipitation of insoluble barium sulfate. A potential role of electrostatically coupled secondary processes that maintain the electroneutrality of the emulsion droplets is discussed. Charging of the GNP metal cores by electron transfer in the presence of the Fe(ii)/Fe(iii) redox couple appears to affect the partitioning of the GNPs between the water droplets and the chloroform phase. Processes have been monitored and studied by optical microscopy, Raman spectroscopy, cryogenic scanning electron microscopy (cryo-SEM) and zeta potential. The shuttle action of the GNPs has further been demonstrated electrochemically in a model system.

Enzymatic activity of lipase-nanoparticle conjugates and the digestion of lipid liquid crystalline assemblies.

Variants of lipase were attached to gold nanoparticles (NPs) and their enzymatic activity was studied. The two bioengineered lipase variants have been prepared with biotin groups attached to different residues on the protein outer surface. The biotinylation was evidenced by denaturing polyacrylamide gel electrophoresis and quantified by the ([2-(4'-hydroxyazobenzene)]benzoic acid spectrophotometric test. NPs of 14 +/- 1 nm diameter coated with thiolated-polyethylene glycol ligands containing controlled proportions of biotin moieties have been prepared and characterized by transmission electron microscopy, UV-vis spectroscopy, small angle neutron scattering, and elemental analysis. These biotin-functionalized NPs were conjugated to lipase using streptavidin as a linker molecule. Enzyme activity assays on the lipase-nanoparticle conjugates show that the lipase loading and activity of the NPs can be controlled by varying the percentage of biotin groups in the particle protecting coat. The lipase-NP conjugates prepared using one variant display higher activity than those prepared using the other variant, demonstrating orientation-dependent enzyme activity. Cryogenic transmission electron microscopy was used to visualize the enzymatic activity of lipase-NP on well-defined lipid substrates. It was found that lipase-coated NPs are able to digest the substrates in a different manner in comparison to the free lipase.

Recyclable molecular trapping and SERS detection in silver-loaded agarose gels with dynamic hot spots.

We describe the design and fabrication of composite agarose gels, highly loaded with silver nanoparticles. Because the gel can collapse upon drying and recover when rehydrated, it can be foreseen as an excellent mechanical molecular trap that additionally gives rise to dynamic hot spots as the network volume decreases and the silver particles get close to each other, thereby generating the high electromagnetic fields that are needed for ultradetection. Additionally, as silver nanoparticles are physically trapped inside the polymer network, analytes can be washed out by dialysis when immersed in a washing solution, so that recycling can be achieved. Finally, the use of SERS for ultradetection of dichlorodiphenyl-trichloroethane (DDT) is reported for the first time, demonstrating the ability of this novel nanocomposite material to reversibly sequester nonconventional SERS analytes.

A multidentate peptide for stabilization and facile bioconjugation of gold nanoparticles.

Gold nanoparticles of two different sizes stabilized by a 15-mer peptide ligand specifically designed for this purpose have been prepared in aqueous solution and characterized by UV-vis spectroscopy and TEM. The presence of the ligand and its binding mode to the particles via its four cystein thiols is evidenced by FTIR and NMR spectroscopy. Biotinylation of the particles via binding to a freely accessible lysine residue is demonstrated.

Electrodeposition of Gold Nanostructures at the Interface of a Pickering Emulsion.

The controlled electrodeposition of nanoparticles at the surface of an emulsion droplet offers enticing possibilities in regards to the formation of intricate structures or fine control over the locus or duration of nanoparticle growth. In this work we develop electrochemical control over the spontaneous reduction of aqueous phase Au(III) by heterogeneous electron transfer from decamethylferrocene present in an emulsion droplet - resulting in the growth of nanoparticles. As gold is a highly effective conduit for the passage of electrical current, even on the nanoscale, the deposition significantly enhances the current response for the single electron transfer of decamethylferrocene when acting as a redox indicator. The nanostructures formed at the surface of the emulsion droplets were imaged by cryo-TEM, providing an insight into the types of structures that may form when stabilised by the interface alone, and how the structures are able to conduct electrons.

Multimodal cell tracking from systemic administration to tumour growth by combining gold nanorods and reporter genes.

Understanding the fate of exogenous cells after implantation is important for clinical applications. Preclinical studies allow imaging of cell location and survival. Labelling with nanoparticles enables high sensitivity detection, but cell division and cell death cause signal dilution and false positives. By contrast, genetic reporter signals are amplified by cell division. Here, we characterise lentivirus-based bi-cistronic reporter gene vectors and silica-coated gold nanorods (GNRs) as synergistic tools for cell labelling and tracking. Co-expression of the bioluminescence reporter luciferase and the optoacoustic reporter near-infrared fluorescent protein iRFP720 enabled cell tracking over time in mice. Multispectral optoacoustic tomography (MSOT) showed immediate biodistribution of GNR-labelled cells after intracardiac injection and successive clearance of GNRs (day 1-15) with high resolution, while optoacoustic iRFP720 detection indicated tumour growth (day 10-40). This multimodal cell tracking approach could be applied widely for cancer and regenerative medicine research to monitor short- and long-term biodistribution, tumour formation and metastasis.

Conserved effects and altered trafficking of Cetuximab antibodies conjugated to gold nanoparticles with precise control of their number and orientation.

Gold nanoparticles (17 nm) have been functionalized with the antiangiogenic monoclonal antibody drug Cetuximab at a well-defined orientation and coverage density of antibodies. Functionalization has been carried out through site-directed chemistry via the selective oxidation of the carbohydrate moiety of antibodies linked to a thiolated hydrazide. A431 tumor cells have been exposed to these conjugates for in vitro evaluation of their effects. In addition to epithelial growth factor receptor blocking, trafficking and signaling alterations were also observed. Thus, the blocking effects of Cetuximab were increased and sustained for a longer time when associated with the nanoparticles. Enhancing antibody therapy effects by decreasing the needed dose and prolonging its effect by avoiding receptor recycling may serve to obtain increased therapeutic benefits for immunotherapy.

Ion Transport across Biological Membranes by Carborane-Capped Gold Nanoparticles.

Carborane-capped gold nanoparticles (Au/carborane NPs, 2-3 nm) can act as artificial ion transporters across biological membranes. The particles themselves are large hydrophobic anions that have the ability to disperse in aqueous media and to partition over both sides of a phospholipid bilayer membrane. Their presence therefore causes a membrane potential that is determined by the relative concentrations of particles on each side of the membrane according to the Nernst equation. The particles tend to adsorb to both sides of the membrane and can flip across if changes in membrane potential require their repartitioning. Such changes can be made either with a potentiostat in an electrochemical cell or by competition with another partitioning ion, for example, potassium in the presence of its specific transporter valinomycin. Carborane-capped gold nanoparticles have a ligand shell full of voids, which stem from the packing of near spherical ligands on a near spherical metal core. These voids are normally filled with sodium or potassium ions, and the charge is overcompensated by excess electrons in the metal core. The anionic particles are therefore able to take up and release a certain payload of cations and to adjust their net charge accordingly. It is demonstrated by potential-dependent fluorescence spectroscopy that polarized phospholipid membranes of vesicles can be depolarized by ion transport mediated by the particles. It is also shown that the particles act as alkali-ion-specific transporters across free-standing membranes under potentiostatic control. Magnesium ions are not transported.

Synthesis of hierarchically porous inorganic-metal site-isolated nanocomposites.

Hierarchically porous inorganic nanocomposites have been synthesized combining interconnected macropores and mesopores with a high loading of site-isolated gold nanoparticles.