Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effecti... more Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effective delivery and regulation of synthetic catalytic systems in cells are challenging due to the complex intracellular environment and catalyst instability. Here, we report the fabrication of protein-sized bioorthogonal nanozymes through the encapsulation of hydrophobic transition metal catalysts into the monolayer of water-soluble gold nanoparticles. The activity of these catalysts can be reversibly controlled by binding a supramolecular cucurbit[7]uril 'gate-keeper' onto the monolayer surface, providing a biomimetic control mechanism that mimics the allosteric regulation of enzymes. The potential of this gated nanozyme for use in imaging and therapeutic applications was demonstrated through triggered cleavage of allylcarbamates for pro-fluorophore activation and propargyl groups for prodrug activation inside living cells.
Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effecti... more Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effective delivery and regulation of synthetic catalytic systems in cells are challenging due to the complex intracellular environment and catalyst instability. Here, we report the fabrication of protein-sized bioorthogonal nanozymes through the encapsulation of hydrophobic transition metal catalysts into the monolayer of water-soluble gold nanoparticles. The activity of these catalysts can be reversibly controlled by binding a supramolecular cucurbit[7]uril 'gate-keeper' onto the monolayer surface, providing a biomimetic control mechanism that mimics the allosteric regulation of enzymes. The potential of this gated nanozyme for use in imaging and therapeutic applications was demonstrated through triggered cleavage of allylcarbamates for pro-fluorophore activation and propargyl groups for prodrug activation inside living cells.
Combination therapy employing proteins and small molecules provides access to synergistic treatme... more Combination therapy employing proteins and small molecules provides access to synergistic treatment strategies. Co-delivery of these two payloads is challenging due to the divergent physico-chemical properties of small molecule and protein cargos. Nanoparticle-stabilized nanocapsules (NPSCs) are promising for combination treatment strategies since they have the potential to deliver small molecule drugs and proteins simultaneously into the cytosol. In this study, we loaded paclitaxel into the hydrophobic core of the NPSC and self-assembled caspase-3 and nanoparticles on the capsule surface. The resulting combination NPSCs showed a higher cytotoxicity of than either of the single agent NPSCs, with synergistic action established using combination index values.
Small (Weinheim an der Bergstrasse, Germany), 2015
A generalized approach to form hybrid organic-inorganic nanocomposite microparticles via the stru... more A generalized approach to form hybrid organic-inorganic nanocomposite microparticles via the structural reorganization of Pickering emulsion templates is described by V. M. Rotello and co-workers on page 1302. Hydrophilic nanoparticles assembled at the oil/water interface react with a hydrophobic polymer, pulling the particles into the oil core. The combined self-assembly and in-situ covalent conjugation strategy provides a facile method to generate robust porous nanocomposites with excellent stability and high capacity.
Low-cost diagnostics for drinking water contamination have the potential to save millions of live... more Low-cost diagnostics for drinking water contamination have the potential to save millions of lives. We report a method that uses inkjet printing to copattern an enzyme-nanoparticle sensor and substrate on a paper-based test strip for rapid detection of bacteria. A colorimetric response is generated on the paper substrate that allows visual detection of contamination without the need for expensive instrumentation. These strips demonstrate a viable nanomanufacturing strategy for low-cost bacterial detection.
Nanostructure-enabled hierarchical assembly holds promise for efficient biocatalyst immobilizatio... more Nanostructure-enabled hierarchical assembly holds promise for efficient biocatalyst immobilization for improved stability in bioprocessing. In this work we demonstrate the use of a hierarchical assembly immobilization strategy to enhance the physicochemical properties and stability of lipase B from Candida antarctica (CaLB). CaLB was complexed with iron oxide nanoparticles followed by interfacial assembly at the surface of an oil-in-water emulsion. Subsequent ring opening polymerization of the oil provided cross-linked microparticles that displayed an increase in catalytic efficiency when compared to the native enzyme and Novozym 435. The hierarchical immobilized enzyme assembly showed no leakage from the support in 50% acetonitrile and could be magnetically recovered across five cycles. Immobilized lipase exhibited enhanced thermal and pH stability, providing 72% activity retention after 24 h at 50 °C (pH 7.0) and 62% activity retention after 24 h at pH 3.0 (30 °C); conditions resulting in complete deactivation of the native lipase.
An effective method for the generation of hybrid organic-inorganic nanocomposite microparticles f... more An effective method for the generation of hybrid organic-inorganic nanocomposite microparticles featuring controlled size and high structural stability is presented. In this process, an oil-in-water Pickering emulsion is formed using hydrophilic amine-functionalized silica nanoparticles. Covalent modification using a hydrophobic maleic anhydride copolymer then alters nanoparticle wettability during crosslinking, causing a core-shell to nanocomposite structural reorganization of the assemblies. The resulting porous nanocomposites maintain discrete microparticle structures and retain payloads in their oil phase even when incubated in competitive solvents such as ethanol.
Methods in molecular biology (Clifton, N.J.), 2013
Gold nanoparticles have been a versatile tool in recent years for the exploration of biological s... more Gold nanoparticles have been a versatile tool in recent years for the exploration of biological systems. However, challenges with purification and adequate surface coverage limit the biocompatibility of gold nanoparticles. Here, we describe a detailed procedure for the synthesis, purification, and functionalization of biologically compatible gold nanoparticles for in vitro and in vivo studies.
Prolonged retention of internalized nanoparticulate systems inside cells improves their efficacy ... more Prolonged retention of internalized nanoparticulate systems inside cells improves their efficacy in imaging, drug delivery, and theranostic applications. Especially, regulating exocytosis of the nanoparticles is a key factor in the fabrication of effective nanocarriers for chemotherapeutic treatments but orthogonal control of exocytosis in the cellular environment is a major challenge. Herein, we present the first example of regulating exocytosis of gold nanoparticles (AuNPs), a model drug carrier, by using a simple host-guest supramolecular system. AuNPs featuring quaternary amine head groups were internalized into the cells through endocytosis. Subsequent in situ treatment of a complementary cucurbit[7]uril (CB[7]) to the amine head groups resulted in the AuNP-CB[7] complexation inside cells, rendering particle assembly. This complexation induced larger particle assemblies that remained sequestered in the endosomes, inhibiting exocytosis of the particles without any observed cytotoxicity.
The use of nanoparticle-stabilized nanocapsules (NPSCs) for the direct cytosolic delivery of siRN... more The use of nanoparticle-stabilized nanocapsules (NPSCs) for the direct cytosolic delivery of siRNA is reported. In this approach, siRNA is complexed with cationic arginine-functionalized gold nanoparticles by electrostatic interactions, with the resulting ensemble self-assembled onto the surface of fatty acid nanodroplets to form a NPSC/siRNA nanocomplex. The complex rapidly delivers siRNA into the cytosol through membrane fusion, a mechanism supported by cellular uptake studies. Using destabilized green fluorescent protein (deGFP) as a target, 90% knockdown was observed in HEK293 cells. Moreover, the delivery of siRNA targeting polo-like kinase 1 (siPLK1) efficiently silenced PLK1 expression in cancer cells with concomitant cytotoxicity.
Traditional cell detection based on biomarkers relies on specific lock-and-key recognition. Howev... more Traditional cell detection based on biomarkers relies on specific lock-and-key recognition. However, many diseases either do not have obvious biomarkers or these markers are currently unknown. Unbiased sensor arrays based on nanoparticles use selective interactions to generate bioprofiles capable of identifying cell type and state. As a result, these arrays can differentiate subtle variations on the cell surfaces, presenting a potential approach for personalized diagnostics.
Drug delivery systems (DDSs) face several challenges including site-specific delivery, stability,... more Drug delivery systems (DDSs) face several challenges including site-specific delivery, stability, and the programmed release of drugs. Engineered nanoparticle (NP) surfaces with responsive moieties can enhance the efficacy of DDSs for in vitro and in vivo systems. This triggering process can be achieved through both endogenous (biologically controlled release) and exogenous (external stimuli controlled release) activation. In this review, we will highlight recent examples of the use of triggered release strategies of engineered nanomaterials for in vitro and in vivo applications.
... S. Agastia, Bradley Duncana, Serkan Eymurb, Murat Tongaa and Vincent M. Rotello*a. a Departme... more ... S. Agastia, Bradley Duncana, Serkan Eymurb, Murat Tongaa and Vincent M. Rotello*a. a Department of Chemistry, University of Massachusetts ... 11, PM Mendes, S. Jacke, K. Critchley, J. Plaza, Y. Chen, K. Nikitin, RE Palmer, JA Preece, SD Evans and D. Fitzmaurice, Langmuir ...
Emulsions stabilized by enzyme-nanoparticle (NP) complexes were used to fabricate robust biocatal... more Emulsions stabilized by enzyme-nanoparticle (NP) complexes were used to fabricate robust biocatalytic scaffolds after core solidification via crosslinking. These biocatalysts feature ease of formation, high retention of enzymatic activity and reusability.
ABSTRACT Portable X-ray Fluorescence (XRF) spectrometry was used to collect elemental data for in... more ABSTRACT Portable X-ray Fluorescence (XRF) spectrometry was used to collect elemental data for in situ hypocaust tiles and floors at an archaeological excavation site near Orvieto, Italy. Data obtained from 2009 to 2011 using three different XRF instruments are compared. Both quantitative results and spectral data were utilized for graphical and chemometric evaluation of the samples' elemental compositions. Despite the different instrumental conditions, changes in data collection protocols, and various data evaluation procedures used with each instrument, conclusions about the archaeological relationships among the tiles remained the same across the three-year study. Utilization of fused data did provide some improvements in differentiation of the materials but at the cost of significantly increased analysis times. The robustness of portable XRF instruments for differentiation of in situ archaeological samples is clearly demonstrated.
... Mary Kate Donais*a, David Georgeb, Bradley Duncanc, Sara M. Wojtasa and AM Daiglea. a Saint A... more ... Mary Kate Donais*a, David Georgeb, Bradley Duncanc, Sara M. Wojtasa and AM Daiglea. a Saint Anselm College ... A portable X-ray fluorescence (XRF) spectrometer and portable Raman spectrophotometer were utilized at the Coriglia, Castel Viscardo excavation site near ...
Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effecti... more Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effective delivery and regulation of synthetic catalytic systems in cells are challenging due to the complex intracellular environment and catalyst instability. Here, we report the fabrication of protein-sized bioorthogonal nanozymes through the encapsulation of hydrophobic transition metal catalysts into the monolayer of water-soluble gold nanoparticles. The activity of these catalysts can be reversibly controlled by binding a supramolecular cucurbit[7]uril 'gate-keeper' onto the monolayer surface, providing a biomimetic control mechanism that mimics the allosteric regulation of enzymes. The potential of this gated nanozyme for use in imaging and therapeutic applications was demonstrated through triggered cleavage of allylcarbamates for pro-fluorophore activation and propargyl groups for prodrug activation inside living cells.
Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effecti... more Bioorthogonal catalysis broadens the functional possibilities of intracellular chemistry. Effective delivery and regulation of synthetic catalytic systems in cells are challenging due to the complex intracellular environment and catalyst instability. Here, we report the fabrication of protein-sized bioorthogonal nanozymes through the encapsulation of hydrophobic transition metal catalysts into the monolayer of water-soluble gold nanoparticles. The activity of these catalysts can be reversibly controlled by binding a supramolecular cucurbit[7]uril 'gate-keeper' onto the monolayer surface, providing a biomimetic control mechanism that mimics the allosteric regulation of enzymes. The potential of this gated nanozyme for use in imaging and therapeutic applications was demonstrated through triggered cleavage of allylcarbamates for pro-fluorophore activation and propargyl groups for prodrug activation inside living cells.
Combination therapy employing proteins and small molecules provides access to synergistic treatme... more Combination therapy employing proteins and small molecules provides access to synergistic treatment strategies. Co-delivery of these two payloads is challenging due to the divergent physico-chemical properties of small molecule and protein cargos. Nanoparticle-stabilized nanocapsules (NPSCs) are promising for combination treatment strategies since they have the potential to deliver small molecule drugs and proteins simultaneously into the cytosol. In this study, we loaded paclitaxel into the hydrophobic core of the NPSC and self-assembled caspase-3 and nanoparticles on the capsule surface. The resulting combination NPSCs showed a higher cytotoxicity of than either of the single agent NPSCs, with synergistic action established using combination index values.
Small (Weinheim an der Bergstrasse, Germany), 2015
A generalized approach to form hybrid organic-inorganic nanocomposite microparticles via the stru... more A generalized approach to form hybrid organic-inorganic nanocomposite microparticles via the structural reorganization of Pickering emulsion templates is described by V. M. Rotello and co-workers on page 1302. Hydrophilic nanoparticles assembled at the oil/water interface react with a hydrophobic polymer, pulling the particles into the oil core. The combined self-assembly and in-situ covalent conjugation strategy provides a facile method to generate robust porous nanocomposites with excellent stability and high capacity.
Low-cost diagnostics for drinking water contamination have the potential to save millions of live... more Low-cost diagnostics for drinking water contamination have the potential to save millions of lives. We report a method that uses inkjet printing to copattern an enzyme-nanoparticle sensor and substrate on a paper-based test strip for rapid detection of bacteria. A colorimetric response is generated on the paper substrate that allows visual detection of contamination without the need for expensive instrumentation. These strips demonstrate a viable nanomanufacturing strategy for low-cost bacterial detection.
Nanostructure-enabled hierarchical assembly holds promise for efficient biocatalyst immobilizatio... more Nanostructure-enabled hierarchical assembly holds promise for efficient biocatalyst immobilization for improved stability in bioprocessing. In this work we demonstrate the use of a hierarchical assembly immobilization strategy to enhance the physicochemical properties and stability of lipase B from Candida antarctica (CaLB). CaLB was complexed with iron oxide nanoparticles followed by interfacial assembly at the surface of an oil-in-water emulsion. Subsequent ring opening polymerization of the oil provided cross-linked microparticles that displayed an increase in catalytic efficiency when compared to the native enzyme and Novozym 435. The hierarchical immobilized enzyme assembly showed no leakage from the support in 50% acetonitrile and could be magnetically recovered across five cycles. Immobilized lipase exhibited enhanced thermal and pH stability, providing 72% activity retention after 24 h at 50 °C (pH 7.0) and 62% activity retention after 24 h at pH 3.0 (30 °C); conditions resulting in complete deactivation of the native lipase.
An effective method for the generation of hybrid organic-inorganic nanocomposite microparticles f... more An effective method for the generation of hybrid organic-inorganic nanocomposite microparticles featuring controlled size and high structural stability is presented. In this process, an oil-in-water Pickering emulsion is formed using hydrophilic amine-functionalized silica nanoparticles. Covalent modification using a hydrophobic maleic anhydride copolymer then alters nanoparticle wettability during crosslinking, causing a core-shell to nanocomposite structural reorganization of the assemblies. The resulting porous nanocomposites maintain discrete microparticle structures and retain payloads in their oil phase even when incubated in competitive solvents such as ethanol.
Methods in molecular biology (Clifton, N.J.), 2013
Gold nanoparticles have been a versatile tool in recent years for the exploration of biological s... more Gold nanoparticles have been a versatile tool in recent years for the exploration of biological systems. However, challenges with purification and adequate surface coverage limit the biocompatibility of gold nanoparticles. Here, we describe a detailed procedure for the synthesis, purification, and functionalization of biologically compatible gold nanoparticles for in vitro and in vivo studies.
Prolonged retention of internalized nanoparticulate systems inside cells improves their efficacy ... more Prolonged retention of internalized nanoparticulate systems inside cells improves their efficacy in imaging, drug delivery, and theranostic applications. Especially, regulating exocytosis of the nanoparticles is a key factor in the fabrication of effective nanocarriers for chemotherapeutic treatments but orthogonal control of exocytosis in the cellular environment is a major challenge. Herein, we present the first example of regulating exocytosis of gold nanoparticles (AuNPs), a model drug carrier, by using a simple host-guest supramolecular system. AuNPs featuring quaternary amine head groups were internalized into the cells through endocytosis. Subsequent in situ treatment of a complementary cucurbit[7]uril (CB[7]) to the amine head groups resulted in the AuNP-CB[7] complexation inside cells, rendering particle assembly. This complexation induced larger particle assemblies that remained sequestered in the endosomes, inhibiting exocytosis of the particles without any observed cytotoxicity.
The use of nanoparticle-stabilized nanocapsules (NPSCs) for the direct cytosolic delivery of siRN... more The use of nanoparticle-stabilized nanocapsules (NPSCs) for the direct cytosolic delivery of siRNA is reported. In this approach, siRNA is complexed with cationic arginine-functionalized gold nanoparticles by electrostatic interactions, with the resulting ensemble self-assembled onto the surface of fatty acid nanodroplets to form a NPSC/siRNA nanocomplex. The complex rapidly delivers siRNA into the cytosol through membrane fusion, a mechanism supported by cellular uptake studies. Using destabilized green fluorescent protein (deGFP) as a target, 90% knockdown was observed in HEK293 cells. Moreover, the delivery of siRNA targeting polo-like kinase 1 (siPLK1) efficiently silenced PLK1 expression in cancer cells with concomitant cytotoxicity.
Traditional cell detection based on biomarkers relies on specific lock-and-key recognition. Howev... more Traditional cell detection based on biomarkers relies on specific lock-and-key recognition. However, many diseases either do not have obvious biomarkers or these markers are currently unknown. Unbiased sensor arrays based on nanoparticles use selective interactions to generate bioprofiles capable of identifying cell type and state. As a result, these arrays can differentiate subtle variations on the cell surfaces, presenting a potential approach for personalized diagnostics.
Drug delivery systems (DDSs) face several challenges including site-specific delivery, stability,... more Drug delivery systems (DDSs) face several challenges including site-specific delivery, stability, and the programmed release of drugs. Engineered nanoparticle (NP) surfaces with responsive moieties can enhance the efficacy of DDSs for in vitro and in vivo systems. This triggering process can be achieved through both endogenous (biologically controlled release) and exogenous (external stimuli controlled release) activation. In this review, we will highlight recent examples of the use of triggered release strategies of engineered nanomaterials for in vitro and in vivo applications.
... S. Agastia, Bradley Duncana, Serkan Eymurb, Murat Tongaa and Vincent M. Rotello*a. a Departme... more ... S. Agastia, Bradley Duncana, Serkan Eymurb, Murat Tongaa and Vincent M. Rotello*a. a Department of Chemistry, University of Massachusetts ... 11, PM Mendes, S. Jacke, K. Critchley, J. Plaza, Y. Chen, K. Nikitin, RE Palmer, JA Preece, SD Evans and D. Fitzmaurice, Langmuir ...
Emulsions stabilized by enzyme-nanoparticle (NP) complexes were used to fabricate robust biocatal... more Emulsions stabilized by enzyme-nanoparticle (NP) complexes were used to fabricate robust biocatalytic scaffolds after core solidification via crosslinking. These biocatalysts feature ease of formation, high retention of enzymatic activity and reusability.
ABSTRACT Portable X-ray Fluorescence (XRF) spectrometry was used to collect elemental data for in... more ABSTRACT Portable X-ray Fluorescence (XRF) spectrometry was used to collect elemental data for in situ hypocaust tiles and floors at an archaeological excavation site near Orvieto, Italy. Data obtained from 2009 to 2011 using three different XRF instruments are compared. Both quantitative results and spectral data were utilized for graphical and chemometric evaluation of the samples' elemental compositions. Despite the different instrumental conditions, changes in data collection protocols, and various data evaluation procedures used with each instrument, conclusions about the archaeological relationships among the tiles remained the same across the three-year study. Utilization of fused data did provide some improvements in differentiation of the materials but at the cost of significantly increased analysis times. The robustness of portable XRF instruments for differentiation of in situ archaeological samples is clearly demonstrated.
... Mary Kate Donais*a, David Georgeb, Bradley Duncanc, Sara M. Wojtasa and AM Daiglea. a Saint A... more ... Mary Kate Donais*a, David Georgeb, Bradley Duncanc, Sara M. Wojtasa and AM Daiglea. a Saint Anselm College ... A portable X-ray fluorescence (XRF) spectrometer and portable Raman spectrophotometer were utilized at the Coriglia, Castel Viscardo excavation site near ...
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