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A self-assembled sensing system able to detect Hg2+ at low nanomolar concentrations is reported that operates through a signal transduction pathway involving multivalent interactions. The analyte causes dimerization of low-affinity ligands, resulting in a complex with a high affinity for a multivalent monolayer-protected gold nanoparticle (AuNP). This complex displaces a quenched fluorescent reporter from the AuNP, resulting in a turn ON of fluorescence. It is shown that the strength of the output signal can be regulated by tuning the multivalent interactions between the complex and the NP. Finally, it is shown that multivalent interactions drive the self-selection of a high-affinity complex from a mixture of low-affinity ligands. [ABSTRACT FROM AUTHOR]

Thisarticle delineates the formation and characterization of differentenzyme–carbon dot conjugates in aqueous medium (pH = 7.0).We used soybean peroxidase (SBP), Chromobacterium viscosum(CV) lipase, trypsin, and cytochrome c(cyt c) for the formation of conjugate either with cationic carbondot (CCD) or anionic carbon dot (ACD) depending on the overall chargeof the protein at pH 7.0. These nanobioconjugates were used to probethe location of enzymes in water-in-oil (w/o) microemulsion. The sizeof the synthesized water-soluble carbon dots were of 2–3 nmwith distinctive emission property. The formation of enzyme/protein–carbondot conjugates in aqueous buffer was confirmed via fluorescence spectroscopyand zeta potential measurement, and the structural alteration of enzyme/proteinwas monitored by circular dichroism spectroscopy. Biocatalytic activitiesof protein/enzymes in conjugation with carbon dots were found to bedecreased in aqueous phosphate buffer (pH 7.0, 25 mM). Interestingly,the catalytic activity of the nanobioconjugates of SBP, CV lipase,and cyt cdid not reduce in cetyltrimethylammoniumbromide (CTAB)-based reverse micelle. It indicates different localizationof carbon dots and the enzymes inside the reverse micelle. The hydrophiliccarbon dots always preferred to be located in the water pool of reversemicelle, and thus, enzyme must be located away from the water pool,which is the interface. However, in case of trypsin–carbondot conjugate, the enzyme activity notably decreased in reverse micellein the presence of carbon dot in a similar way that was observed inwater. This implies that trypsin and carbon dots both must be locatedat the same place, which is the water pool of reverse micelle. Carbondot induced deactivation was not observed for those enzymes whichstay away from the water pool and localized at the interfacial domainwhile deactivation is observed for those enzymes which reside at thewater pool. Thus, the location of enzymes in the microdomain of w/omicroemulsion can be predicted by comparing the activity profile ofenzyme–carbon dot conjugate in water and w/o microemulsion. [ABSTRACT FROM AUTHOR]

Although cellular transport machinery is mostly ATP-driven and ATPase-dependent, there has been a recent surge in understanding colloidal transport processes relying on a nonspecific physical interaction with biologically significant small molecules. Herein, we probe the phoretic behavior of a biocolloid [composed of a Zn(II)-coordinated metallomicelle and enzymes horseradish peroxidase (HRP) and glucose oxidase (GOx)] when exposed to a concentration gradient of ATP under microfluidic conditions. Simultaneously, we demonstrate that an ATP-independent oxidative biocatalytic product formation zone can be modulated in the presence of a (glucose + ATP) gradient. We report that both directionality and extent of transport can be tuned by changing the concentration of the ATP gradient. This diffusiophoretic mobility of a submicrometer biocolloidal object for the spatial transposition of a biocatalytic zone signifies the ATP-mediated functional transportation without the involvement of ATPase. Additionally, the ability to analyze colloidal transport in microfluidic channels using an enzymatic fluorescent product-forming reaction could be a new nanobiotechnological tool for understanding transport and spatial catalytic patterning processes. We believe that this result will inspire further studies for the realization of elusive biological transport processes and target-specific delivery vehicles, considering the omnipresence of the ATP-gradient across the cell.

The nascent field of microscale flow chemistry focuses on harnessing flowing fluids to optimize chemical reactions in microchambers and establish new routes for chemical synthesis. With enzymes and other catalysts anchored to the surface of microchambers, the catalytic reactions can act as pumps and propel the fluids through the containers. Hence, the flows not only affect the catalytic reactions, but these reactions also affect the flows. Understanding this dynamic interplay is vital to enhancing the accuracy and utility of flow technology. Through experiments and simulation, we design a system of three different enzymes, immobilized in separate gels, on the surface of a microchamber; with the appropriate reactants in the solution, each enzyme-filled gel acts as a pump. The system also exploits a reaction cascade that controls the temporal interactions between two pumps. With three pumps in a triangular arrangement, the spatio-temporal interactions among the chemical reactions become highly coordinated and produce well-defined fluid streams, which transport chemicals and form a fluidic "circuit". The circuit layout and flow direction of each constituent stream can be controlled through the number and placement of the gels and the types of catalysts localized in the gels. These studies provide a new route for forming self-organizing and bifurcating fluids that can yield fundamental insight into nonequilibrium, dynamical systems. Because the flows and fluidic circuits are generated by internal chemical reactions, the fluids can autonomously transport cargo to specific locations in the device. Hence, the findings also provide guidelines to facilitate further automation of microfluidic devices.

A self-assembled sensing system able to detect Hg(2+) at low nanomolar concentrations is reported that operates through a signal transduction pathway involving multivalent interactions. The analyte causes dimerization of low-affinity ligands, resulting in a complex with a high affinity for a multivalent monolayer-protected gold nanoparticle (AuNP). This complex displaces a quenched fluorescent reporter from the AuNP, resulting in a turn ON of fluorescence. It is shown that the strength of the output signal can be regulated by tuning the multivalent interactions between the complex and the NP. Finally, it is shown that multivalent interactions drive the self-selection of a high-affinity complex from a mixture of low-affinity ligands.

The present work reports covalent functionalization of single-walled carbon nanotubes (f-SWNTs) to introduce hydrophilicity to the otherwise amphiphobic nanotubes. The charge and spacer length of the functional moiety were varied by using quaternized ethylene diamine, 6-aminocaproate, quaternized (ethylenedioxy)bis(ethylamine), and a poly(ethylene glycol) (PEG) unit (f-SWNT-1 to f-SWNT-4, respectively). These f-SWNTs with varying degrees of hydrophilicity were incorporated within cetyltrimethyl ammonium bromide (CTAB) reverse micelles to develop stable self-assembled nanohybrids. An optimum hydrophilicity on the SWNT surface led to interfacial localization of f-SWNTs resulting in the augmentation of space at the interface. A surface-active enzyme, lipase, localized at this enhanced interface of f-SWNT-containing CTAB reverse micelles exhibited significant activation (2.5-fold) compared to that in the absence of the nanoconstructs. This improvement in lipase activity was mainly due to the smooth occupancy of lipase and also presumably because of the increase in the concentrations of both substrate and the enzyme at the augmented interface. Interestingly, the f-SWNTs that activate lipase in reverse micelles deactivate the same enzyme in water. The dispersion of f-SWNTs in water and its matching integration at the interface of reverse micelles were confirmed through transmission electron microscopic (TEM) investigations. The interfacial localization of these nanoconstructs was also established from the distinct fluorescence behavior of a hydrophobic fluorescent probe, fluorescein isothiocyanate (FITC), adsorbed onto the f-SWNT surface. In concurrence with the observed lipase activity, the corresponding changes in the enzyme conformation within f-SWNTs integrated reverse micelle as well as in aqueous medium were studied by circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy.