Utilization of Spontaneous Alkyne-Gold Self-Assembly Chemistry as an Alternative Method for Fabricating Electrochemical Aptamer-Based Sensors.

Electrochemical aptamer-based (E-AB) sensors are a promising class of biosensors which use structure-switching redox-labeled oligonucleotides (aptamers) codeposited with passivating alkanethiol monolayers on electrode surfaces to specifically bind and detect target analytes. Signaling in E-AB sensors is an outcome of aptamer conformational changes upon target binding, with the sequence of the aptamer imparting specificity toward the analyte of interest. The change in conformation translates to a change in electron transfer between the redox label attached to the aptamer and the underlying electrode and is related to analyte concentration, allowing specific electrochemical detection of nonelectroactive analytes.

Single Voltammetric Sweep Calibration-Free Interrogation of Electrochemical Aptamer-Based Sensors Employing Continuous Square Wave Voltammetry.

Continuous square wave voltammetry (cSWV) is a technique that enables the continuous collection of current data (at 100 kHz) to maximize the information content obtainable from a single voltammetric sweep. This data collection procedure results in the generation of multiple voltammograms corresponding to different effective square wave frequencies. The application of cSWV brings significant benefits to electrochemical aptamer-based (E-AB) sensors.

Tuning the Probe-Bilayer Architecture of Silver Nanoneedle-based Ion Channel Probes.

Ion channel probes, as one of the ion channel platforms, provide an appealing opportunity to perform localized detection with a high precision level. These probes come basically in two classes: glass and metal. While the glass-based probes showed the potential to be employed for molecular sensing and chemical imaging, these probes still suffer from limited resolution and lack of control over protein insertion.

Controlling the Collision Type and Frequency of Single Pt Nanoparticles at Chemically Modified Gold Electrodes.

Studying the electrochemical response of single nanoparticles at an electrode surface gives insight into the dynamic and stochastic processes that occur at the electrode interface. Herein, we investigated single platinum nanoparticle collision dynamics and type (elastic vs inelastic) at gold electrode surfaces modified with self-assembled monolayers (SAMs) of varying terminal chemistries. Collision events are measured via the faradaic current from catalytic reactions at the Pt surface.

Perspective-Assessing Electrochemical, Aptamer-Based Sensors for Dynamic Monitoring of Cellular Signaling.

Electrochemical, aptamer-based (E-AB) sensors provide a generalizable strategy to quantitatively detect a variety of targets including small molecules and proteins. The key signaling attributes of E-AB sensors (sensitivity, selectivity, specificity, and reagentless and dynamic sensing ability) make them well suited to monitor dynamic processes in complex environments. A key bioanalytical challenge that could benefit from the detection capabilities of E-AB sensors is that of cell signaling, which involves the release of molecular messengers into the extracellular space.

Microscale, Electrochemical, Aptamer-Based Sensors for Enhanced Small-Molecule Detection at Millisecond Time Scales.

Microscale electrodes offer the advantages of increased mass transport rates, high sensitivity, and rapid measurement capabilities. Fabricating electrochemical aptamer-based (E-AB) sensors on these electrode platforms opens new applications to chemical and biological sensing but has remained challenging due to low signal-to-noise ratios and monolayer instability. In this article, we report the development and characterization of E-AB sensors on a gold microelectrode platform (∼500 nm radius).

Development of an Electrochemical, Aptamer-Based Sensor for Dynamic Detection of Neuropeptide Y.

The ability to monitor dynamic changes in neuropeptide Y (NPY) levels in complex environments can have an impact on many fields, including neuroscience and immunology. Here, we describe the development of an electrochemical, aptamer-based (E-AB) sensor for the dynamic (reversible) measurement of physiologically relevant (nanomolar) concentrations of neuropeptide Y. The E-AB sensors are fabricated using a previously described 80 nucleotide aptamer reported to specifically bind NPY with a binding affinity = 0.

Continuous Square Wave Voltammetry for High Information Content Interrogation of Conformation Switching Sensors.

Square wave voltammetry (SWV) is a voltammetric technique for measuring Faradaic current while minimizing contributions from non-Faradaic processes. In square wave voltammetry, the potential waveform applied to a working electrode and the current sampling protocols followed are designed to minimize contributions from non-Faradaic processes (i.e.

Maintaining Single-Channel Recordings on a Silver Nanoneedle through Probe Design and Feedback Tip Positioning Control.

Ion channel proteins showed great promise in the field of nanopore sensing and molecular flux imaging applications due to the atomic-level precision of the pore size and a high signal-to-noise ratio. More specifically, ion channel probes, where the protein channels are integrated at the end of a solid probe, can achieve highly localized detection. Metal probe materials such as gold and silver have been developed to support lipid bilayers and enable the use of smaller probes, or nanoneedles, compared to more traditional glass micropipette ion channel probes.

Solution-Phase Electrochemical Aptamer-Based Sensors.

Electrochemical aptamer-based sensors (EABs) using self-assembled monolayers on gold working-electrodes have now been in-vivo demonstrated for multiple-analytes, demonstrating their sensitivity and specificity even in a continuous sensing format. However, longevity has been demonstrated for only 24 hours and sensitivity has been challenging for highly dilute analytes (nM regime). A novel approach is reported here using electrochemical aptamer-based sensing that is not covalently-bound to a gold-working electrode but where aptamers are freely mobile in solution.

Recent advances in ion-channel probes for nanopore sensing: Insights into the probe architectures.

This review introduces the recent advances in the nanopore sensing platform, ion channel probes (ICPs), with a particular focus on the different probe design (2011-2022). The use of ion channel proteins has emerged in different applications to understand the dynamics of many biological processes and characterize or detect biomolecules. The development of utilizing protein channels in nanopore sensing has led to diverse platforms in which the ion channels, or biological nanopores, can be embedded in a lipid membrane.

Observing Real-Time Formation of Self-Assembled Monolayers on Polycrystalline Gold Surfaces with Scanning Electrochemical Cell Microscopy.

Self-assembled monolayers (SAMs) of alkanethiols on gold have become a central focus of controllable surface chemistry because they can be easily formed from the solution phase and characterized using various techniques. Understanding the formation processes occurring at a nanoscale level is crucial to form defect-free SAMs for tailored applications in bio- and nanotechnology. Although many reports study and characterize SAMs after they are formed on gold surfaces, typical methods have not extensively studied the SAM formation process at the nanoscale.

Effects of Nucleic Acid Structural Heterogeneity on the Electrochemistry of Tethered Redox Molecules.

The cation condensation-induced collapse of electrode-bound nucleic acids and the resulting change in the electrochemical signal is a useful tool to predict the structure and redox probe location of heterogeneous structures of surface-tethered DNA probes─a common architecture employed in the development of electrochemical sensors. In this paper, we measure the faradaic current of an appended redox molecule at the 3' position of the nucleic acid using cyclic voltammetry before and after nucleic acid collapse for various nucleic acid architectures and heterogeneous mixtures on the same electrode surface. The voltammetric peak current change with collapse correlates with the proximity of the redox molecules from the surface.

3-D printed microfluidics for rapid prototyping and testing of electrochemical, aptamer-based sensor devices under flow conditions.

We demonstrate the ability to rapidly prototype and fabricate an epoxy-embedded electrode platform and microfluidic device suitable for using electrochemical biosensors under flow conditions. We utilize three-dimensional (3-D) printing to rapidly prototype molds to fabricate epoxy-embedded electrodes in addition to molds for rapid prototyping of PDMS microfluidic components. We characterize the bare gold epoxy-embedded electrodes using ferricyanide as a redox indicator and then characterize the performance of an adenosine triphosphate (ATP) specific electrochemical, aptamer-based (E-AB) sensor.

Sensitive Electrochemical Detection of Microcystin-LR in Water Samples Via Target-Induced Displacement of Aptamer Associated [Ru(NH)].

In this study, we demonstrate the successful development of an electrochemical aptamer-based sensor for point-of-use detection and quantification of the highly potent microcystin-LR (MC-LR) in water. The sensor uses hexaammineruthenium(III) chloride ([Ru(NH)]) as redox mediator, because of the ability of the positively charged (3+) molecule to associate with the phosphate backbone of the nucleic acids. We quantitatively measure the target-induced displacement of aptamer associated, or surface confined, [Ru(NH)] in the presence of MC-LR.

Nucleic Acid Identity, Structure, and Flexibility Affect the Electrochemical Signal of Tethered Redox Molecules upon Biopolymer Collapse.

We demonstrate that cation condensation can induce the collapse of surface-bound nucleic acids and that the electrochemical signal from a tethered redox molecule (methylene blue) upon collapse reports on nucleic acid identity, structure, and flexibility. Furthermore, the correlation of the electrochemical signal and structure is consistent with theoretical considerations of nucleic acid collapse. Changes in solution dielectric permittivity or the concentration of trivalent cations cause the structure of nucleic acids to become more compact due to an increase in attractive electrostatic interactions between the charged biopolymer backbone and multivalent ions in the solution.

Silver Nanoneedle Probes Enable Sustained DC Current, Single-Channel Resistive Pulse Nanopore Sensing.

Resistive pulse sensing using ion channel proteins (biological nanopores) has been evolving as a single-molecule approach to detect small biomolecules owing to atomically precise pore size reproducibility, high signal-to-noise ratio, and molecular selectivity. The incorporation of biological nanopores in sensing platforms requires a stable lipid membrane that can be formed by a variety of methods such as the painting method and droplet-based techniques. However, these methods are limited by the fragility of the unsupported bilayer or the need for specific microdevices.

Repetitive drug releases from light-activatable micron-sized liposomes.

In this work, a novel light activatable micron-sized liposomal drug carrier that has a unique capability to release drug repetitively in proportion to the cycle number of short irradiation (5 s) of near-infrared (NIR) pulsed laser is reported. We synthesized methotrexate (MTX)-loaded liposomes based on a modified reverse-phase evaporation method. Gold nanorods (AuNR) were attached to the liposomal surfaces, enabling the liposomes to release drug under short NIR irradiation via the photothermal effect.

Electrochemical Affinity Assays/Sensors: Brief History and Current Status.

The advent of electrochemical affinity assays and sensors evolved from pioneering efforts in the 1970s to broaden the field of analytes accessible to the selective and sensitive performance of electrochemical detection. The foundation of electrochemical affinity assays/sensors is the specific capture of an analyte by an affinity element and the subsequent transduction of this event into a measurable signal. This review briefly covers the early development of affinity assays and then focuses on advances in the past decade.

Effects of Experimental Conditions on the Signaling Fidelity of Impedance-Based Nucleic Acid Sensors.

Electrochemical impedance spectroscopy (EIS), an extremely sensitive analytical technique, is a widely used signal transduction method for the electrochemical detection of target analytes in a broad range of applications. The use of nucleic acids (aptamers) for sequence-specific or molecular detection in electrochemical biosensor development has been extensive, and the field continues to grow. Although nucleic acid-based sensors using EIS offer exceptional sensitivity, signal fidelity is often linked to the physical and chemical properties of the electrode-solution interface.

Use of Electrocatalysis for Differentiating DNA Polymorphisms and Enhancing the Sensitivity of Electrochemical Nucleic Acid-Based Sensors with Covalent Redox Tags-Part II.

Single-nucleotide polymorphisms (SNPs), insertion/deletion (indel) polymorphisms, and DNA methylation are the most frequent types of genetic variations. As such, DNA polymorphisms play significant roles in genetic mapping and diagnostics. Thus, analytical methods enabling DNA polymorphism detection will provide an invaluable means for early disease diagnosis.

Electrocatalytic Mechanism for Improving Sensitivity and Specificity of Electrochemical Nucleic Acid-Based Sensors with Covalent Redox Tags-Part I.

The design and development of advanced electrocatalysis have been extensively explored for efficient energy conversion and electrochemical biosensing. Both ferricyanide (Fe(CN)) and methylene blue (MB) have been widely used in the development of electrochemical biosensing strategies. However, the electrocatalytic mechanism between nucleic acid-tethered MB and Fe(CN) remains unexplored.

Analysis of Membrane Protein Deinsertion-Associated Currents with Nanoneedle-Supported Bilayers to Discover Pore Formation Mechanisms.

Analysis of the pore formation mechanisms of biological nanopores can provide insight into pore-forming peptide-induced diseases and into the characterization of nanopores employed in sensing methods. Evaluation of pore formation mechanisms is typically performed using microscopy including atomic force microscopy, transmission electron microscopy, as well as electrically via channel current measurements using a patch-clamp amplifier. However, due to the relatively low temporal resolution of the above-mentioned microscopy techniques and the low analysis accuracy of the channel current measurements, new analytical methods are required.

Recessed Ag/AgCl Microelectrode-Supported Lipid Bilayer for Nanopore Sensing.

Biological nanopores reconstituted into supported lipid bilayer membranes are widely used as a platform for stochastic nanopore sensing with the ability to detect single molecules including, for example, single-stranded DNA (ssDNA) and miRNA. A main thrust in this area of research has been to improve overall bilayer stability and ease of measurements. These improvements are achieved through a variety of clever strategies including droplet-based techniques; however, they typically require specific microfabrication techniques to prepare devices or special manipulation techniques for microdroplets.

Effect of Laser Irradiation on Reversibility and Drug Release of Light-Activatable Drug-Encapsulated Liposomes.

Although several studies have demonstrated repetitive drug release using light-activatable liposomes, inconsistent drug release at each activation limits widespread usage. Here, we report reversible plasmonic material-coated encapsulated liposomes for proportional controlled delivery of methotrexate (MTX), which is a common drug for cancer and autoimmune diseases, using repetitive laser irradiation. Our results suggest a proportional increase in total drug release after repetitive laser irradiation.