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Single-domain antibodies, known as nanobodies, have great potential as biorecognition elements for sensors because of their small size, affinity, specificity, and robustness. However, facile and efficient methods of nanobody immobilization are sought that retain their maximum functionality. Herein, we describe the direct immobilization of nanobodies on gold sensors by exploiting a modified cysteine strategically positioned at the C-terminal end of the nanobody. The experimental data based on secondary ion mass spectrometry, circular dichroism, and surface plasmon resonance, taken together with a detailed computational work (molecular dynamics simulations), support the formation of stable and well-oriented nanobody monolayers. Furthermore, the nanobody structure and activity is preserved, wherein the nanobody is immobilized at a high density (approximately 1 nanobody per 13 nm2). The strategy for the spontaneous nanobody self-assembly is simple and effective and possesses exceptional potential to be used in numerous sensing platforms, ranging from clinical diagnosis to environmental monitoring.

Sialic acid recognition remains an interesting and challenging target in molecular receptor design. Herein, we report a series of benzoboroxole-based receptors in which cationic hydrogen-bond donors have been introduced and shown to promote multipoint sialic acid recognition. One striking feature revealed by these receptors is that the carboxylate sialic acid residue is the primary binding determinant for recognition by benzoboroxole, in which the presence of charge-reinforced hydrogen bonds results in enhanced selectivity for sialic acid over other carbohydrates and a 4.5-fold increase in affinity. These findings open up wide possibilities for benzoboroxole-based receptors use in life science research, biotechnology, and diagnostics.

The ability to design surfaces with reversible, high-affinity protein binding sites represents a significant step forward in the advancement of analytical methods for diverse biochemical and biomedical applications. Herein, we report a dynamic supramolecular strategy to directly assemble proteins on surfaces based on multivalent host–guest interactions. The host–guest interactions are achieved by one-step nanofabrication of a well-oriented β-cyclodextrin host-derived self-assembled monolayer on gold (β-CD-SAM) that forms specific inclusion complexes with hydrophobic amino acids located on the surface of the protein. Cytochrome c, insulin, α-chymotrypsin, and RNase A are used as model guest proteins. Surface plasmon resonance and static time-of-flight secondary ion mass spectrometry studies demonstrate that all four proteins interact with the β-CD-SAM in a specific manner via the hydrophobic amino acids on the surface of the protein. The β-CD-SAMs bind the proteins with high nanomolar to single-digit micromolar dissociation constants (KD). Importantly, while the proteins can be captured with high affinity, their release from the surface can be achieved under very mild conditions. Our results expose the great advantages of using a supramolecular approach for controlling protein immobilization, in which the strategy described herein provides unprecedented opportunities to create advanced bioanalytic and biosensor technologies.

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Many cell membrane functions emerge from the lateral presentation of membrane receptors. The link between the nanoscale organization of the receptors and ligand binding remains, however, mostly unclear. In this work, we applied surface molecular imprinting and utilized the phase behavior of lipid bilayers to create platforms that recapitulate the lateral organization of membrane receptors at the nanoscale. We used liposomes decorated with amphiphilic boronic acids that commonly serve as synthetic saccharide receptors and generated three lateral modes of receptor presentation─random distribution, nanoclustering, and receptor crowding─and studied their interaction with saccharides. In comparison to liposomes with randomly dispersed receptors, surface-imprinted liposomes resulted in more than a 5-fold increase in avidity. Quantifying the binding affinity and cooperativity proved that the boost was mediated by the formation of the nanoclusters rather than a local increase in the receptor concentration. In contrast, receptor crowding, despite the presence of increased local receptor concentrations, prevented multivalent oligosaccharide binding due to steric effects. The findings demonstrate the significance of nanometric aspects of receptor presentation and generation of multivalent ligands including artificial lectins for the sensitive and specific detection of glycans.

Herein, we report a highly sensitive electrocatalytic sensor-cell construct that can electrochemically communicate with the internal environment of immune cells (e.g., macrophages) via the selective monitoring of a particular reactive oxygen species (ROS), hydrogen peroxide. The sensor, which is based on vertically aligned single-walled carbon nanotubes functionalized with an osmium electrocatalyst, enabled the unprecedented detection of a local intracellular “pulse” of ROS on a short second time scale in response to bacterial endotoxin (lipopolysaccharide-LPS) stimulation. Our studies have shown that this initial pulse of ROS is dependent on NADPH oxidase (NOX) and toll like receptor 4 (TLR4). The results suggest that bacteria can induce a rapid intracellular pulse of ROS in macrophages that initiates the classical innate immune response of these cells to infection.

Under the exerted external electric field (Eex), significant polarization effects are demonstrated in the switching process of positively charged oligopeptide chains on the gold surface by using the polarizable force-field-based molecular dynamics (MD) simulations. The changes of electrostatic environment during the conformational switching are described by calling density functional theory (DFT) calculations of atomic charges or fragment-centered dipole moments. The charge-variable polarizable force field (Q-POL) model shows a good agreement with the experimental observations of both the open-circuit state (OC) without Eexand extended/bent conformation (ON/OFF) with upward/downward Eex, while the nonpolarizable force field (Non-POL) with the fixed atomic partial charges fails to reproduce experiments for the ON state. The charged oligopeptide chains (with each residue furnished with a positive charge) challenge the application of the coarse-graining polarizable force field based on the fragment dipoles (Dfrag-POL). Through tracing the dynamics of charge centers of both counterions and oligopeptide chains in the switching process, a qualitative picture is depicted for understanding the polarization phenomena in the dielectric monolayer in OC/ON/OFF states. With the applied electric field, the rearrangement of ions leads to the induced internal E-field, Ein, in the direction opposite to Eex. The values of total electric field, Etotal, and the interfacial potential Φxwere evaluated. The relationship between the applied electric potential and the interfacial potential is built for the switching OC/ON/OFF states. The analysis of the induced internal E-field and interfacial potential may shed light on the further modification of theoretical models to better understand the electrical-induced switching mechanism.

The rational design of materials with tailored properties is of paramount importance for a wide variety of biological, medical, electronic and optical applications. Here we report molecular level control over the spatial distribution of functional groups on surfaces utilizing self-assembled monolayers (SAMs) of pH-switchable surface-appended pseudorotaxanes. The supramolecular systems were constructed from a poly(aryl ether) dendron-containing a dibenzo[24]crown-8 (DB24C8) macrocycle and a thiol ligand-containing a dibenzylammonium recognition site and a fluorine end group. The dendron establishes the space (dendritic effect) that each pseudorotaxane occupies on the SAM. Following SAM formation, the dendron is released from the surface by switching off the noncovalent interactions upon pH stimulation, generating surface materials with tailored physical and chemical properties.

Understanding how bacteria adhere to a surface is a critical step in the development of novel materials and coatings to prevent bacteria forming biofilms. Here, surface plasmon resonance (SPR) spectroscopy, in combination with self‐assembled monolayers (SAMs) that have different backbone structures and/or functional groups, is used for the first time to study the initial stages of bacterial adhesion to surfaces (i.e., initial interaction of cells with a surface, a process governed by van der Waals, electrostatic, and hydrophobic interactions). The work highlights SPR spectroscopy as a powerful and unique approach to probe bacterial adhesion in real time. SPR spectral data reveal different kinetics of adhesion for the interaction of two marine bacterial species (Marinobacter hydrocarbonoclasticusand Cobetia marina)to a range of organosulfur SAMs. Furthermore, the extent of adhesion is dependent on the backbone structures and functional groups of the SAMs. The role of extracellular polymeric substances (EPS) in bacterial adhesion is also investigated. Pre‐conditioning experiments with cell‐free culture supernatants, containing planktonic EPS, allow quantification of the amount adsorbed onto surfaces and directly account for the impact of EPS adsorption on bacterial adhesion in the assay. While the physicochemical characteristics of the surfaces play a significant role in determining bacterial cell adhesion for low levels of conditioning by planktonic EPS, greater levels of conditioning by EPS reduce the difference between surfaces.

The ability to regulate biomolecular interactions on surfaces driven by an external stimuli is of great theoretical interest and practical impact in the biomedical and biotechnology fields. Herein, a new class of responsive surfaces that rely on electro‐switchable peptides to control biomolecular interactions on gold surfaces is presented. This system is based upon the conformational switching of positively charged oligolysine peptides that are tethered to a gold surface, such that bioactive molecular moieties (biotin) incorporated on the oligolysines can be reversibly exposed (bio‐active state) or concealed (bio‐inactive state) on demand, as a function of surface potential. The dynamics of switching the biological properties is studied by observing the binding events between biotin and fluorescently labeled NeutrAvidin. Fluorescence microscope images and surface plasmon resonance spectral data clearly reveal opposite binding behaviors when +0.3 V or −0.4 V vs. SCE are applied to the surface. High fluorescence intensities are observed for an applied positive potential, while minimal fluorescence is detected for an applied negative potential. Surface plasmon resonance spectroscopy (SPR) results provided further evidence that NeutrAvidin binding to the surface is controlled by the applied potential. A large SPR response is observed when a positive potential is applied on the surface, while a negative applied potential induces over 90% reduction in NeutrAvidin binding.

Amine monofunctional gold nanoparticles (1-AuNPs) were synthesized by employing a solid-supported technique and pH-switchable pseudorotaxane formation. Purification was repeatedly facilitated using crown ether peripherally coated superparamagnetic iron oxide microspheres to yield the monofunctional gold nanoparticles in excellent yield. The product and its related intermediate superstructures were characterized by IR and X-ray photoelectron spectroscopies. Novel supramolecular dimers and trimers were prepared by titrating the 1-AuNPs with bisDB24C8 and trisDB24C8 at different ratios. UV/visible absorption spectroscopic analyses of the supramolecular dimer and trimer solutions, which were formed by mixing their separate components in different ratios, indicated the gradual appearance of two distinct plasmonic resonance bands at 620 and ∼700 nm. Furthermore, TEM images of the dimers revealed a significant amount of dimer pairs on the surface, while the TEM images of the trimers demonstrated the presence of both dimers and trimers. The trimers appeared as triangular or near-linear shapes.

The dynamic nature of micellar nanostructures is employed to form a self-assembled Förster resonance energy transfer (FRET) nanoplatform for enhanced sensing of DNA. The platform consists of lipid oligonucleotide FRET probes incorporated into micellar scaffolds, where single recognition events result in fusion and fission of DNA mixed micelles, triggering the fluorescence response of multiple rather than a single FRET pair. In comparison to conventional FRET substrates where a single donor interacts with a single acceptor, the micellar multiplex FRET system showed ∼20- and ∼3-fold enhancements in the limit of detection and FRET efficiency, respectively. This supramolecular signal amplification approach could potentially be used to improve FRET-based diagnostic assays of nucleic acid and non-DNA based targets.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.