Targeting small molecule drugs to T cells with antibody-directed cell-penetrating gold nanoparticles

We sought to develop a nanoparticle vehicle that could efficiently deliver small molecule drugs to target lymphocyte populations. The synthesized amphiphilic organic ligand-protected gold nanoparticles (amph-NPs) were capable of sequestering large payloads of small molecule drugs within hydrophobic pockets of their ligand shells. These particles exhibit membrane-penetrating activity in mammalian cells, and thus enhanced uptake of a small molecule TGF-β inhibitor in T cells in cell culture. By conjugating amph-NPs with targeting antibodies or camelid-derived nanobodies, the particles' cell-penetrating properties could be temporarily suppressed, allowing targeted uptake in specific lymphocyte subpopulations. Degradation of the protein targeting moieties following particle endocytosis allowed the NPs to recover their cell-penetrating activity in situ to enter the cytoplasm of T cells. In vivo, targeted amph-NPs showed 40-fold enhanced uptake in CD8+ T cells relative to untargeted particles, and delivery of TGF-β inhibitor-loaded particles to T cells enhanced their cytokine polyfunctionality in a cancer vaccine model. Thus, this system provides a facile approach to concentrate small molecule compounds in target lymphocyte populations of interest for immunotherapy in cancer and other diseases.Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (Contract W911NF-13-D-0001)Melanoma Research AllianceNational Cancer Institute (U.S.) (David H. Koch Institute for Integrative Cancer Research at MIT. (Support (Core) Grant P30-CA14051)National Institutes of Health (U.S.) (Grant CA174795)National Institutes of Health (U.S.) (Grant CA172164)Horizon 2020 Framework Programme (European Commission). FutureNanoNeeds Projec

Near-field excitation and near-field detection of propagating surface plasmon polaritons on Au waveguide structures

The propagation of surface plasmon polaritons guided along Au metal waveguides fabricated by electron-beam lithography is experimentally investigated using simultaneous near-field excitation and detection of plasmon-polariton modes localized at the air/Au interface. The directly measured propagation characteristics of surface plasmon-polaritons agree well with simulation results obtained using full-vector calculations and the analytic dispersion of asymmetric plasmonic waveguides for thin Au films. Our results demonstrate that near-field excitation/detection schemes are well suited for direct imaging and characterization of propagating surface plasmon-fields bound to thin-film metal layers, and can be used for fast and reliable characterization of plasmonic waveguide elements and nanodevices

Thermodynamic Study of the Reactivity of the Two Topological Point Defects Present in Mixed Self-Assembled Monolayers on Gold Nanoparticles

When a mixed self-assembled monolayer forms on the curved surface of a gold nanoparticle two opposed point defects must form for topological reasons. Here we present a thermodynamic study proving that these defects are more reactive than other sites on the nanoparticle, and demonstrate that the energy of the reaction depends on the ligand molecules

Effects of Surface Compositional and Structural Heterogeneity on Nanoparticle-Protein Interactions: Different Protein Configurations

As nanoparticles (NPs) enter into biological systems, they are immediately exposed to a variety and concentration of proteins. The physicochemical interactions between proteins and NPs are influenced by the surface properties of the NPs. To identify the effects of NP surface heterogeneity, the interactions between bovine serum albumin (BSA) and gold NPs (AuNPs) with similar chemical composition but different surface structures were investigated. Different interaction modes and BSA conformations were studied by dynamic light scattering, circular dichroism spectroscopy, fluorescence quenching and isothermal titration calorimetry (ITC). Depending on the surface structure of AuNPs, BSA seems to adopt either a "side-on" or an "end-on" conformation on AuNPs. ITC demonstrated that the adsorption of BSA onto AuNPs with randomly distributed polar and nonpolar groups was primarily driven by electrostatic interaction, and all BSA were adsorbed in the same process. The adsorption of BSA onto AuNPs covered with alternating domains of polar and nonpolar groups was a combination of different interactions. Overall, the results of this study point to the potential for utilizing nanoscale manipulation of NP surfaces to control the resulting NP-protein interactions

A simple atomic force microscopy method for the visualization of polar and non-polar parts in thin organic films

Here we present a scanning probe microscopy method that allows for the identification of regions of different polarity (i.e. hydrophilicity) in thin organic films. This technique is based on the analysis of the difference between phase images generated at different applied bias voltages in tapping-mode atomic force microscopy. We show that, without any chemical modification of the microscope tip, it is possible to investigate surface properties of complex macromolecular layers, yielding new insight into the functional properties of the photosynthetic electron transport macromolecular complex, Photosystem I

A scalable synthesis of highly stable and water dispersible Ag-44(SR)(30) nanoclusters

We report the synthesis of atomically monodisperse thiol-protected silver nanoclusters [Ag-44(SR)(30)](m), (SR = 5-mercapto-2-nitrobenzoic acid) in which the product nanocluster is highly stable in contrast to previous preparation methods. The method is one-pot, scalable, and produces nanoclusters that are stable in aqueous solution for at least 9 months at room temperature under ambient conditions, with very little degradation to their unique UV-Vis optical absorption spectrum. The composition, size, and monodispersity were determined by electrospray ionization mass spectrometry and analytical ultracentrifugation. The produced nanoclusters are likely to be in a superatom charge-state of m = 4-, due to the fact that their optical absorption spectrum shares most of the unique features of the intense and broadly absorbing nanoparticles identified as [Ag-44(SR)(30)](4-) by Harkness et al. (Nanoscale, 2012, 4, 4269). A protocol to transfer the nanoclusters to organic solvents is also described. Using the disperse nanoclusters in organic media, we fabricated solid-state films of [Ag-44(SR)(30)](m) that retained all the distinct features of the optical absorption spectrum of the nanoclusters in solution. The films were studied by X-ray diffraction and photoelectron spectroscopy in order to investigate their crystallinity, atomic composition and valence band structure. The stability, scalability, and the film fabrication method demonstrated in this work pave the way towards the crystallization of [Ag-44(SR)(30)](m) and its full structural determination by single crystal X-ray diffraction. Moreover, due to their unique and attractive optical properties with multiple optical transitions, we anticipate these clusters to find practical applications in light-harvesting, such as photovoltaics and photocatalysis, which have been hindered so far by the instability of previous generations of the cluster

Calcium-triggered fusion of lipid membranes is enabled by amphiphilic nanoparticles

Lipid membrane fusion is an essential process for a number of critical biological functions. The overall process is thermodynamically favorable but faces multiple kinetic barriers along the way. Inspired by nature's engineered proteins such as SNAP receptor [soluble N-ethylmale-imide-sensitive factor-attachment protein receptor (SNARE)] complexes or viral fusogenic proteins that actively promote the development of membrane proximity, nucleation of a stalk, and triggered expansion of the fusion pore, here we introduce a synthetic fusogen that can modulate membrane fusion and equivalently prime lipid membranes for calcium-triggered fusion. Our fusogen consists of a gold nanoparticle functionalized with an amphiphilic monolayer of alkanethiol ligands that had previously been shown to fuse with lipid bilayers. While previous efforts to develop synthetic fusogens have only replicated the initial steps of the fusion cascade, we use molecular simulations and complementary experimental techniques to demonstrate that these nanoparticles can induce the formation of a lipid stalk and also drive its expansion into a fusion pore upon the addition of excess calcium. These results have important implications in general understanding of stimuli-triggered fusion and the development of synthetic fusogens for biomedical applications.U.S. Department of Energy (Contract DE-FG02-97ER25308)National Science Foundation (Contract TG-DMR130042)U. S. Army Research Office (Contract W911NF-13-D-0001