Electrochemical Aptamer Sensor Research Articles

Synthesis of SnC@Au@Apta by electrospinning as an electrochemical sensor for detection of tetracycline

A highly efficient electrochemical aptamer sensor for the detection of tetracycline (TC) was prepared by using SnC@Au@Apta. Metal tin has good electrochemical activity and high conductivity. It is often used as an electrochemical sensing material. The nanofibers prepared by electrospinning machine make the metal distribution more uniform, not easy to agglomerate, and have a certain porosity, which can improve the sensitivity of sensor detection. Carbonization further enhances conductivity. The gold nanoparticles (AuNPs) on the surface of SnC nanomaterials improve the electrochemical detection performance, and also act as the binding site of the TC aptamer, which is stably combined with the thiol group at the end of the TC aptamer. The TC aptamer specifically binds to TC to detect TC in the sample. The electrochemical performance of SnC@Au@Apta was evaluated by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Under the optimal conditions, the detection range of SnC@Au@Apta is wide (0.001–100 μM), the detection limit is low (0.83 nM), and it has excellent selectivity, stability and reproducibility. In addition, SnC@Au@Apta can be used to detect TC in milk samples.

Nanopillar array-based electrochemical aptamer sensor for STX sensitivity detection.

Saxitoxin (STX) is a cyanotoxin with high toxicity, and therefore, there is an urgent need to develop a facile detection method for STX. In this study, an ordered nanopillar array-based electrochemical aptasensor was fabricated for the high-performance detection of STX. The anti-STX aptamer with methylene blue (MB) incorporated at the 3'-end (MB-Apt) was immobilized at the surface of an Au@PAN nanopillar array electrode and used as the recognition element. The proposed aptasensor demonstrated highly sensitive and selective STX detection because of synergistic catalysis effects of MB and ordered nanopillar arrays along with the selection of MB-Apt. The nanopillar array-based electrochemical aptasensor exhibited high sensitivity over a wide linear concentration range of 1 pM-3 nM with a linear regression equation of ΔI (μA) = 28.0 + 6.9 × log[STX] (R2 = 0.98079) and 3-100 nM with a linear regression equation of ΔI (μA) = 10.7 + 43.4 × log[STX] (R2 = 0.98772), where R is the correlation coefficient. In addition, the limit of detection (LOD) was as low as 1 pM. Furthermore, the designed aptasensor demonstrated excellent selectivity toward STX, preventing interference from neo-STX, okadaic acid, and common metal ions. The presented orderly nanopillar array-based strategy to develop an electrochemical aptasensor for STX detection offers a promising method for developing high-performance electrochemical sensors, and the presented aptasensor should find useful application in the detection of shellfish poison.

Detection of AFB1 in corn by MXene paper‐based unlabeled aptasensor

AbstractIn this work, a paper‐based electrochemical aptamer sensor was developed for the detection of aflatoxin B1 (AFB1) using a combination of MXene–Ti3C2Tx and nucleic acid aptamers. The prepared single‐layer or few‐layer MXene suspension is suction‐filtered onto MXene paper, which is cut to prepare MXene electrodes. To accomplish AFB1 specific detection, an amino‐labeled AFB1 aptamer is mounted on the surface of the carboxy‐functionalized MXene electrode. When AFB1 is present, it particularly binds to the aptamer to form a 3D structure, reducing the efficiency of electron transmission on the sensor surface. The difference in impedance signal change at the electrode/electrolyte interface is used to quantify AFB1. The results indicated that the detection range is 0.05–100 ng/mL, the detection limit is 0.04 ng/mL, and the recovery rate of AFB1 in corn samples is 97.8%–111.52% with the optimal detection conditions. The MXene paper‐based label‐free aptasensor is versatile and can detect different targets by simply swapping out the aptamers of different targets. The sensor also has a wide range of applications in food analysis and environmental testing.Practical applicationsA paper‐based electrochemical aptamer sensor was developed to detect aflatoxin B1 using a combination of MXene–Ti3C2Tx and nucleic acid aptamers.The design is based on the preparation of MXene electrodes by pumping and filtering monolayer or multilayer MXene suspensions onto MXene paper and cutting.The MXene paper‐based label‐free aptamer sensor was designed to be versatile, allowing the detection of different targets by simply replacing the aptamer with one from a different targets.

A structure-switching electrochemical aptamer sensor for mercury ions based on an ordered assembled gold nanorods-modified electrode

In this paper, an electrochemical aptamer sensor with high sensitivity and selectivity was proposed for the detection of mercury ion (Hg2+). The ordered assembled gold nanorods-modified screen-printed electrode (AuNRs/SPE) was used to improve the sensor with higher sensitivity and data reproducibility. The sensor was designed based on the specific recognition of Hg2+ with aptamer to form T-Hg2+-T binding structure. Two aptamer chains were applied in the designed sensor, aptamer S1 with terminal thiolate group was self-assembled on the AuNRs/SPE via Au–S bond, and complementary S2 labeled with ferrocene (Fc) would generate a strong redox electrical signal. The addition of Hg2+ would mediate the folding of the T-base of S1 to form a hairpin structure, resulting in the dissociation of the Fc-labeled S2 from the sensing system and a significant reduction in the redox current. The electrochemical properties of the sensor were further investigated using cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The effects of aptamer reaction time, concentration, and target incubation time on the sensor response were also investigated. The results showed that the sensor had a sensitive response to Hg2+ at concentrations ranging from 1 pM to 1 μM, with a detection limit of 0.3 pM. Furthermore, the proposed sensor was applied in the determination of Hg2+ in real samples, demonstrating its potential practical value.

Portable electrochemical aptasensor for highly sensitive detection of 3,3′,4,4′-tetrachlorobiphenyl

Aptamer-based electrochemical sensors are frequently used as independent, surface-functionalized, passive electrodes. However, their sensitivity and detection limits become limited, particularly when the electrode area is reduced to facilitate miniaturization. A mobile phone-based microfluidic electrochemical aptamer sensing platform for 3,3′,4,4′-tetrachlorobiphenyl (PCB77) detection was developed in this work. This aptamer sensor utilized Exonuclease I (Exo I) and DNA/AuNPs/horseradish peroxidase (DNA/AuNPs/HRP) nanoprobes as a merged signal amplification method, which resulted in an increase in the electrochemical sensing performance. Sensitive detection of PCB77 was accomplished by functionalizing the hierarchically structured Au@MoS2/CNTs/GO modified working/sensing electrode with the specific aptamer. The aptamer sensor was tested with different concentrations of PCB77 within the microfluidic platform. Afterward, the differential pulse voltammograms were recorded using a wireless integrated circuit device. Subsequently, the collected data was transmitted to a smartphone using Bluetooth communication. A detection limit of 0.0085 ng/L was obtained for PCB77 detection, with a detection range from 0.1 to 1000 ng/L. In addition, the detection of PCB77 in spiked water samples validated the possibility of using this aptamer sensor in a real environment, and the aptamer sensor demonstrated high selectivity in distinguishing PCB77 from other potential interfering species. The merging of electrochemical aptamer sensors with purposefully engineered microfluidic and integrated devices in this study is a novel and promising method that provides a dependable platform for on-site applications.

An innovative label-free electrochemical aptamer sensor: utilizing Ti3C2Tx/MoS2/Au NPs for accurate interleukin-6 detection

In the medical field, changes in interleukin-6 (IL-6) concentration serve as essential biomarkers for monitoring and diagnosing various conditions, including acute inflammatory responses such as those seen in trauma and burns, and chronic illnesses like cancer. This paper detailed a label-free electrochemical aptamer sensor designed for IL-6 quantification. A composite material consisting of Ti3C2Tx and MoS2 was successfully synthesized to fabricate this sensor. The synergistic effect of MoS2's catalytic action on hydrogen peroxide (H2O2), used as a signalling marker, when combined with the exceptional conductivity and large specific surface area of Ti3C2Tx, not only enables an increased loading of MoS2 but also significantly boosts the electrochemical response. The in situ-reduced Au NPs provided stable immobilization sites for DNA aptamers (DNAapt) and facilitated electron transfer, ensuring accurate IL-6 recognition. Under optimal conditions, the aptamer sensor exhibited a wide linear range (5 pg/mL to 100 ng/mL) and a low limit of detection (LOD) of 2.9 pg/mL. Its sensing performance in human serum samples highlights its potential as a promising clinical analysis tool.

Thymine-Hg2+-Thymine strategy in MOF-based electrochemical aptamer sensor for PAEs detection

Phthalate acid esters (PAEs), known food contaminants, can lead to reproductive problems, respiratory diseases, childhood obesity, and neuropsychological disorders. This work aims to develop an electrochemical aptamer sensor for the detection of DAP and its analogs. To achieve specific recognition ability for PAEs, we have introduced the thymidine-Hg2+-thymidine strategy for the first time in an electrochemical aptamer sensor, incorporating metal–organic frameworks (MOFs) as the carrier for the aptamer. Additionally, nano signal amplification technology was applied by immobilizing MXene on the electrode to intensify the sensor’s current signal. In the presence of PAEs, these molecules specifically bound to the aptamers, resulting in competitive release of Hg2+ and aptamers connected to Hg2+, subsequently reducing current signaling. Under optimal experimental conditions, the designed sensor achieved a linear range from 1.65 × 10−5 mg/mL to 3 × 10−3 mg/mL and an extremely low detection limit of 8.94 × 10−6 mg/mL. Furthermore, it exhibited good reliability and practicality in detecting actual samples.

Research Progress in the Detection of Aflatoxin B1 Based on Aptamers

Abstract: Aflatoxin B1 is highly toxic, mutagenic, teratogenic, and carcinogenic and is a class I carcinogen. Peanuts, cotton, and corn may be affected by AFB1 during cultivation, which can seriously jeopardize human health. Developing a simple, sensitive, and selective method for detecting AFB1 is imminent. Aptamers are obtained through in vitro screening of ligands by single-stranded oligonucleotides (DNA or RNA) through exponential enrichment (SELEX) technology. As emerging highly selective recognition molecules, they have the advantages of strong affinity, good stability, and strong specificity. Because it does not have the function of signal conversion, it cannot produce physicochemical signals that can be detected in the process of specific binding with target molecules, so it is necessary to convert the process of specific binding of aptamers to target molecules into a process of easily detectable physicochemical signal changes. According to different conversion methods, aptamer biosensors are divided into electrochemical aptamer sensors, fluorescent aptamer sensors, colorimetric aptamer sensors, surface Raman-enhanced aptamer sensors, and so on. Herein, the recent progress and application of aflatoxin B1 detection by nucleic acid aptamer biosensors based on the above signals are reviewed, and the future development prospects and challenges of this kind of biosensor are summarized.

A Sensitive Electrochemical Aptamer Sensor for Clenbuterol Detection Based on Au NPs/Fe3O4–NH2/GN Nanocomposite

Chronic ingestion of food containing clenbuterol (CLB) in the human body can cause acute poisoning. Thus, it is essential to develop a sensitive CLB detection way. Here, a novel electrochemical aptamer sensor based on aptamer/gold nanoparticles/amino magnetic ferric oxide/graphene nanosheets (aptamer/Au NPs/Fe3O4–NH2/GN) nanocomposite has been developed. The Fe3O4–NH2 nanoparticles were self-assembled on GN, and then Fe3O4–NH2/GN nanocomposite was coated on the glassy carbon electrode (GCE) surface. Afterward, Au NPs were electrodeposited on Fe3O4–NH2/GN nanocomposite to fix the aptamer by the Au–S bonds. When CLB was added to the solution, the aptamer will specifically recognize CLB, causing current changes. Under the optimized conditions, the sensor shows better electrochemical performance (linear range is [Formula: see text][Formula: see text]M to [Formula: see text][Formula: see text]M, the detection limit is [Formula: see text][Formula: see text]M ([Formula: see text])). It has been successfully applied to the detection of CLB in actual samples. The results show that the aptamer sensor has good selectivity, stability and repeatability, which will provide a novel idea for more sensitive and quicker CLB detection.

Dual-mode colorimetric electrochemical aptamer sensor based on Au@Pd bimetallic nanoparticles for the detection of Pb(II) in edible mushrooms

Dual-mode colorimetric electrochemical aptamer sensor based on Au@Pd bimetallic nanoparticles for the detection of Pb(II) in edible mushrooms

Electrochemical aptamer sensor based on AgNPs@PDANSs and "sandwich" structure guidance for the detection of tobramycin in water samples.

In this study, an ultrasensitive detection platform for tobramycin (TOB) was developed, featuring a "sandwich" structure guided by AgNCs@PDANSs and Thi-AuNCs@ZnONSs. To address the issue of large background current peak signals in tagless sensors, Thi-AuNCs@ZnONSs composites were synthesized as signal tags. Zinc oxide nanosheets (ZnONSs) served as the loading agent, and AuNCs with the electroactive molecule Thi acted as carriers. Furthermore, AgNPs@PDANSs nanocomposites, possessing excellent electrical conductivity and large specific surface areas, were prepared as substrate materials for the modified electrodes. A "sandwich" structure strategy was also introduced to enhance the accuracy of the electrochemical aptasensor. This strategy, utilizing a dual sequence for target labeling and capture, yielded higher sensitivity and simplified the sensor construction compared to methods employing a single sequence. Under optimal conditions, the detection limit for TOB was established at 1.41 pM, with a detection range of 0.05-5000 nM. The aptasensor was effectively applied in the detection of TOB in tap and lake water, demonstrating outstanding reproducibility, selectivity, and stability. These results may serve as a reference for environmental TOB detection.

An electrochemical aptamer sensor for rapid quantification sulfadoxine based on synergistic signal amplification of indole and MWCNTs and its electrooxidation mechanism

Unreasonable disposal of sulfadoxine (SDX) causing food safety and eventually result water pollution, and shows great significance of detection. A sensitive unit Aptamer (Apt) was adopted to construct an electrochemical sensor BSA/Apt/indole/MWCNTs/GCE with the synergistic amplification of multi-walled carbon nanotubes (MWCNTs) and indole for the detection of SDX. Optimization of the conditions allowed the detection of SDX shows a wide linear range from 0.10 to 1000.00 µmol/L by differential pulse voltammetry (DPV). The LOD and sensitivity were determined to be 0.033 µmol/L and 0.29 µA/(µmol/L·cm²), respectively. The recovery rate of SDX in pork and lake water samples range 98.57 %−104.93 %, 93.6 %−106.2 %, respectively. Both the relative standard deviation (RSD) was lower than 7.40 %, which proved that the sensor has good practicability. In addition, DFT calculation was established that a restricted of B3LYP and a basis set of 6–31 G, the electrooxidation of SDX involves two-electron and one-proton transfer.

Shared hairpin structure electrochemical aptasensor based on ZrO2@Ni/Co-MOFs@AuNPs for dual-target detection of Cd2+ and S.aureus

Heavy metals and foodborne pathogens have become key pollutants that harm aquatic products, seriously endangering food safety and human health. Therefore, in this study, a ratiometric electrochemical aptamer sensor with a novel signal amplification strategy was constructed using ZrO2 as the substrate and Ni/Co-MOFs@AuNPs cubic nanomaterials to increase the specific surface area to simultaneously detect Cd2+ and S.aureus. The shared hairpin complementary chains modified with ferropene (Fc) could simultaneously bind to the Cd2+ and S.aureus aptamers modified with methylene blue (MB) and anthrquinone-2-carboxylic acid (AQ). The ratios of MB and AQ to Fc were used to realize the qualitative and quantitative detection of Cd2+ and S.aureus. Under the optimal conditions, the electrochemical aptamer sensor for Cd2+ and S.aureus was in the range of 1.00 × 10−2-1.00 × 104 ng/L and 1.00 × 101-1.00 × 107 CFU/mL, and the lowest detection limit was 1.91 × 10−4 ng/L and 1.37 CFU/mL, respectively. The aptasensor constructed in this paper can realize the simultaneous detection of Cd2+ and S.aureus in scallop and fish, providing a new detection method for the simultaneous detection of a variety of aquatic products pollutants.

Truncated Electrochemical Aptasensor with Enhanced Antifouling Capability for Highly Sensitive Serotonin Detection.

Accurate determination of serotonin (ST) provides insight into neurological processes and enables applications in clinical diagnostics of brain diseases. Herein, we present an electrochemical aptasensor based on truncated DNA aptamers and a polyethylene glycol (PEG) molecule-functionalized sensing interface for highly sensitive and selective ST detection. The truncated aptamers have a small size and adopt a stable stem-loop configuration, which improves the accessibility of the aptamer for the analyte and enhances the sensitivity of the aptasensor. Upon target binding, these aptamers perform a conformational change, leading to a variation in the Faraday current of the redox tag, which was recorded by square wave voltammetry (SWV). Using PEG as blocking molecules minimizes nonspecific adsorption of other interfering molecules and thus endows an enhanced antifouling ability. The proposed electrochemical aptamer sensor showed a wide range of detection lasting from 0.1 nM to 1000 nM with a low limit of detection of 0.14 nM. Owing to the unique properties of aptamer receptors, the aptasensor also exhibits high selectivity and stability. Furthermore, with the reduced unspecific adsorption, assaying of ST in human serum and artificial cerebrospinal fluid (aCSF) showed excellent performance. The reported strategy of utilizing antifouling PEG describes a novel approach to building antifouling aptasensors and holds great potential for neurochemical investigations and clinical diagnosis.

Study on a nano-porous gold/polyamidoamine (NPG/PAMAM)-based electrochemical aptamer biosensor for the detection of ochratoxin a in the red wine

In this study, a novel electrochemical aptamer sensor for detecting ochratoxin A (OTA) was constructed. First, a gold-copper alloy film was prepared via electrochemical deposition, and copper was selectively dissolved in constant potential mode for obtaining the nano-porous gold modified screen-printed carbon electrodes (NPG/SPCE). Then, 2-mercaptoethylamine was dropped on the NPG/SPCE surface and Au-S covalent bonds were formed for immobilizing the metal. Glutaraldehyde as cross-linking agent was added, which resulted in immobilization and attachment of PAMAM to the 2-mercaptoethylamine through the dehydration condensation reaction. During the preparation process, the nano-porous gold and PAMAM-modified layers were characterized by SEM, XRD, and IR spectroscopy, respectively. The characterization results showed that the nano-porous gold and PAMAM composite films were successfully modified. Finally, the OTA aptamer was cross-linked with PAMAM by glutaraldehyde to complete construction of the Apt/PAMAM/NPG/SPCE sensor. The electrochemical performance of this sensor was tested in ochratoxin A solutions with the DPV method. The results showed that the sensor’s reproducibility, stability, and specificity were good. The spiked recoveries in red wine ranged from 99.65%∼101.6%, with a linear range of 0.5 ng/mL∼20 ng/mL and a minimum detection limit of 0.141 ng/mL. Thus, the novel biosensor may provide a promising tool for the trace detection of OTA.

Homogeneous Electrochemical Aptamer Sensor Based on Two-Dimensional Nanocomposite Probe and Nanochannel Modified Electrode for Sensitive Detection of Carcinoembryonic Antigen.

A rapid and convenient homogeneous aptamer sensor with high sensitivity is highly desirable for the electrochemical detection of tumor biomarkers. In this work, a homogeneous electrochemical aptamer sensor is demonstrated based on a two-dimensional (2D) nanocomposite probe and nanochannel modified electrode, which can realize sensitive detection of carcinoembryonic antigen (CEA). Using π-π stacking and electrostatic interaction, CEA aptamer (Apt) and cationic redox probe (hexaammineruthenium(III), Ru(NH3)63+) are co-loaded on graphite oxide (GO), leading to a 2D nanocomposite probe (Ru(NH3)63+/Apt@GO). Vertically ordered mesoporous silica-nanochannel film (VMSF) is easily grown on the supporting indium tin oxide (ITO) electrode (VMSF/ITO) using the electrochemical assisted self-assembly (EASA) method within 10 s. The ultrasmall nanochannels of VMSF exhibits electrostatic enrichment towards Ru(NH3)63+ and size exclusion towards 2D material. When CEA is added in the Ru(NH3)63+/Apt@GO solution, DNA aptamer recognizes and binds to CEA and Ru(NH3)63+ releases to the solution, which can be enriched and detected by VMSF/ITO electrodes. Based on this mechanism, CEA can be an electrochemical detection ranging from 60 fg/mL to 100 ng/mL with a limit of detection (LOD) of 14 fg/mL. Detection of CEA in human serum is also realized. The constructed homogeneous detection system does not require the fixation of a recognitive aptamer on the electrode surface or magnetic separation before detection, demonstrating potential applications in rapid, convenient and sensitive electrochemical sensing of tumor biomarkers.

Nanohybrid of antimonene@Ti3C2Tx-based electrochemical aptasensor for lead detection

Lead ions (Pb2+), as one of many common heavy metallic environmental pollutants, can cause serious side-effects and result in chronic poisoning to people's health, so it is highly significant to monitor Pb2+ efficiently and sensitively. Here, we proposed an antimonene@Ti3C2Tx nanohybrid-based electrochemical aptamer sensor (aptasensor) for high sensitive Pb2+ determination. The sensing platform of nanohybrid was synthesized by ultrasonication, possessing the advantages of both antimonene and Ti3C2Tx, which not only can vastly enlarge the sensing signal of the proposed aptasensor, but also greatly simplified its manufacturing flow, because antimonene can strongly interact with aptamer through noncovalently bound. The surface morphology and microarchitecture of the nanohybrid were perused by several methods such as scanning electron microscope (SEM), energy-dispersive X-ray mapping spectroscopy (EDS), transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), and atomic force microscope (AFM). Under optimal empirical conditions, the proposed aptasensor exhibited a wide linear correlation of the current signals with the logarithm of CPb2+ (Log CPb2+) over the span from 1 × 10−12 to 1 × 10−7 M and provided a trace discernment limit of 3.3 × 10−13 M. Moreover, the constructed aptasensor displayed superior repeatability, great consistency, eminent selectivity, and beneficial reproducibility, implying its extreme potential application for water quality control and the environmental monitoring of Pb2+.

A review of advances in aptamer-based cell detection technology.

Since cells are the basic structural and functional units of organisms, the detection or quantitation of cells is one of the most common basic problems in life science research. The established cell detection techniques mainly include fluorescent dye labeling, colorimetric assay, and lateral flow assay, all of which employ antibodies as cell recognition elements. However, the widespread application of the established methods generally dependent on antibodies is limited, because the preparation of antibodies is complicated and time-consuming, and unrecoverable denaturation is prone to occur with antibodies. By contrast, aptamers that are generally selected through the systematic evolution of ligands by exponential enrichment can avoid the disadvantages of antibodies due to their controllable synthesis, thermostability, and long shelf life, etc. Accordingly, aptamers may serve as novel molecular recognition elements like antibodies in combination with various techniques for cell detection. This paper reviews the developed aptamer-based cell detection methods, mainly including aptamer-fluorescent labeling, aptamer-isothermal amplification assay, electrochemical aptamer sensor, aptamer-based lateral flow analysis, and aptamer-colorimetric assay. The principles, advantages, progress of application in cell detection and future development trend of these methods were specially discussed. Overall, different assays are suitable for different detection purposes, and the development of more accurate, economical, efficient, and rapid aptamer-based cell detection methods is always on the road in the future. This review is expected to provide a reference for achieving efficient and accurate detection of cells as well as improving the usefulness of aptamers in the field of analytical applications.

Week-Long Operation of Electrochemical Aptamer Sensors: New Insights into Self-Assembled Monolayer Degradation Mechanisms and Solutions for Stability in Serum at Body Temperature.

Conventional wisdom suggests that widely utilized self-assembled alkylthiolate monolayers on gold are too unstable to last more than several days when exposed to complex fluids such as raw serum at body temperature. Demonstrated here is that these monolayers can not only last at least 1 week under such harsh conditions but that significant applied value can be captured for continuous electrochemical aptamer biosensors. Electrochemical aptamer biosensors provide an ideal tool to investigate monolayer degradation, as aptamer sensors require a tightly packed monolayer to preserve sensor signal vs background current and readily reveal fouling by albumin and other solutes when operating in biofluids. Week-long operation in serum at 37 °C is achieved by (1) increasing van der Waals interactions between adjacent monolayer molecules to increase the activation energy required for desorption, (2) optimizing electrochemical measurement to decrease both alkylthiolate oxidation and electric-field-induced desorption, and (3) mitigating fouling using protective zwitterionic membranes and zwitterion-based blocking layers with antifouling properties. This work further proposes origins and mechanisms of monolayer degradation in a logical stepwise manner that was previously unobservable over multiday time scales. Several of the observed results are surprising, revealing that short-term improvements to sensor longevity (i.e., hours) actually increase sensor degradation in the longer term (i.e., days). The results and underlying insights on mechanisms not only push forward fundamental understanding of stability for self-assembled monolayers but also demonstrate an important milestone for continuous electrochemical aptamer biosensors.

Continuous molecular monitoring of human dermal interstitial fluid with microneedle-enabled electrochemical aptamer sensors.

The ability to continually collect diagnostic information from the body during daily activity has revolutionized the monitoring of health and disease. Much of this monitoring, however, has been of physical "vital signs", with the monitoring of molecular markers having been limited to glucose, primarily due to the lack of other medically relevant molecules for which continuous measurements are possible in bodily fluids. Electrochemical aptamer sensors, however, have a recent history of successful in vivo demonstrations in rat animal models. Herein, we present the first report of real-time human molecular data collected using such sensors, successfully demonstrating their ability to measure the concentration of phenylalanine in dermal interstitial fluid after an oral bolus. To achieve this, we used a device that employs three hollow microneedles to couple the interstitial fluid to an ex vivo, phenylalanine-detecting sensor. The resulting architecture achieves good precision over the physiological concentration range and clinically relevant, 20 min lag times. By also demonstrating 90 days dry room-temperature shelf storage, the reported work also reaches another important milestone in moving such sensors to the clinic. While the devices demonstrated are not without remaining challenges, the results at minimum provide a simple method by which aptamer sensors can be quickly moved into human subjects for testing.