Single-Walled Carbon Nanotubes as Enhancing Substrates for PNA-Based Amperometric Genosensors

A new amperometric sandwich-format genosensor has been implemented on single-walled carbon nanotubes screen printed electrodes (SWCNT-SPEs) and compared in terms of performance with analogous genoassays developed using the same methodology on non-nanostructured glassy carbon platforms (GC-SPE). The working principle of the genosensors is based on the covalent immobilization of Peptide Nucleic Acid (PNA) capture probes (CP) on the electrode surface, carried out through the carboxylic functions present on SWCNT-SPEs (carboxylated SWCNT) or electrochemically induced on GC-SPEs. The sequence of the CP was complementary to a 20-mer portion of the target DNA; a second biotin-tagged PNA signalling probe (SP), with sequence complementary to a different contiguous portion of the target DNA, was used to obtain a sandwich hybrid with an Alkaline Phosphatase-streptavidin conjugate (ALP-Strp). Comparison of the responses obtained from the SWCNT-SPEs with those produced from the non-nanostructured substrates evidenced the remarkable enhancement effect given by the nanostructured electrode platforms, achieved both in terms of loading capability of PNA probes and amplification of the electron transfer phenomena exploited for the signal transduction, giving rise to more than four-fold higher sensitivity when using SWCNT-SPEs. The nanostructured substrate allowed to reach limit of detection (LOD) of 71 pM and limit of quantitation (LOQ) of 256 pM, while the corresponding values obtained with GC-SPEs were 430 pM and 1.43 nM, respectively

Design of specific nucleic acid‐based biosensors for protein binding activity

Nucleic acid-based biosensors for the detection of specific proteins combine the typical programmability of synthetic DNA systems with artificially controlled DNA-protein communication. The high-affinity interaction between a target protein and a specific ligand, such as an aptamer sequence, or a double stranded DNA domain, or a small peptide, is paired with a nature-mimicking molecular mechanism allowing for probing, processing, and translating protein binding activity into a measurable signal. In this Review, two main strategies developed in the context of protein-responsive nucleic acid-based biosensors are discussed. One is the design of proximity-based assays harnessing the spatial colocalization of functional probes within the volume of a multivalent protein. The other is the engineering of dynamic DNA structures that undergo a controlled conformational or structural change upon protein binding. Examples of applications from optical and electrochemical detection of antibodies in biofluids to fluorescence imaging of transcription factors in living cells are reported, and suggestions along with possible future directions in the field are discussed

A Wi-Fi cloud-based portable potentiostat for electrochemical biosensors

The measurement of the analyte concentration in electrochemical biosensors traditionally requires costly laboratory equipment to obtain accurate results. Innovative portable solutions have recently been proposed, but usually, they lean on personal computers (PCs) or smartphones for data elaboration and they exhibit poor resolution or portability and proprietary software. This paper presents a low-cost portable system, assembling an ad hoc -designed analog front end (AFE) and a development board equipped with a system on chip integrating a microcontroller and a Wi-Fi network processor. The wireless module enables the transmission of measurements directly to a cloud service for sharing device outcome with users (physicians, caregivers, and so on). In doing so, the system does not require neither the customized software nor other devices involved in data acquisition. Furthermore, when any Internet connection is lost, the data are stored on board for subsequent transmission when a Wi-Fi connection is available. The noise output voltage spectrum has been characterized. Since the designed device is intended to be battery-powered to enhance portability, investigations about battery lifetime were carried out. Finally, data acquired with a conventional benchtop Autolab PGSTAT-204 electrochemical workstation are compared with the outcome of our developed device to validate the effectiveness of our proposal. To this end, we selected ferri/ferrocyanide as redox probe, obtaining the calibration curves for both the platforms. The final outcomes are shown to be feasible, accurate, and repeatable

Controlling Dynamic DNA Reactions at the Surface of Single-Walled Carbon Nanotube Electrodes to Design Hybridization Platforms with a Specific Amperometric Readout

: Carbon nanotube (CNT)-based electrodes are cheap, highly performing, and robust platforms for the fabrication of electrochemical sensors. Engineering programmable DNA nanotechnologies on the CNT surface can support the construction of new electrochemical DNA sensors providing an amperometric output in response to biomolecular recognition. This is a significant challenge, since it requires gaining control of specific hybridization processes and functional DNA systems at the interface, while limiting DNA physisorption on the electrode surface, which contributes to nonspecific signal. In this study, we provide design rules to program dynamic DNA structures at the surface of single-walled carbon nanotubes electrodes, showing that specific DNA interactions can be monitored through measurement of the current signal provided by redox-tagged DNA strands. We propose the use of pyrene as a backfilling agent to reduce nonspecific adsorption of reporter DNA strands and demonstrate the controlled formation of DNA duplexes on the electrode surface, which we then apply in the design and conduction of programmable DNA strand displacement reactions. Expanding on this aspect, we report the development of novel amperometric hybridization platforms based on artificial DNA structures templated by the small molecule melamine. These platforms enable dynamic strand exchange reactions orthogonal to conventional toehold-mediated strand displacement and may support new strategies in electrochemical sensing of biomolecular targets, combining the physicochemical properties of nanostructured carbon-based materials with programmable nucleic acid hybridization

A Folding-Based Electrochemical Aptasensor for the Single-Step Detection of the SARS-CoV-2 Spike Protein

Efficient and timely testing has taken center stage in the management, control, and monitoring of the current COVID-19 pandemic. Simple, rapid, cost-effective diagnostics are needed that can complement current polymerase chain reaction-based methods and lateral flow immunoassays. Here, we report the development of an electrochemical sensing platform based on single-walled carbon nanotube screen-printed electrodes (SWCNT-SPEs) functionalized with a redox-tagged DNA aptamer that specifically binds to the receptor binding domain of the SARS-CoV-2 spike protein S1 subunit. Single-step, reagentless detection of the S1 protein is achieved through a binding-induced, concentration-dependent folding of the DNA aptamer that reduces the efficiency of the electron transfer process between the redox tag and the electrode surface and causes a suppression of the resulting amperometric signal. This aptasensor is specific for the target S1 protein with a dissociation constant (K-D) value of 43 +/- 4 nM and a limit of detection of 7 nM. We demonstrate that the target S1 protein can be detected both in a buffer solution and in an artificial viral transport medium widely used for the collection of nasopharyngeal swabs, and that no cross-reactivity is observed in the presence of different, non-target viral proteins. We expect that this SWCNT-SPE-based format of electrochemical aptasensor will prove useful for the detection of other protein targets for which nucleic acid aptamer ligands are made available

IoT and Biosensors: A Smart PortablePotentiostat With AdvancedCloud-Enabled Features

Recent advances in Internet-of-Things technology have opened the doors to new scenariosfor biosensor applications. Flexibility, portability, and remote control and access are of utmost importanceto move these devices to people’s homes or in a Point-of-Care context and rapidly share the results withusers and their physicians. In this paper, an innovative portable device for both quantitative and semi-quantitative electrochemical analysis is presented. This device can operate autonomously without the needof relying on other devices (e.g., PC, tablets, or smartphones) thanks to built-in Wi-Fi connectivity. Thedeveloped hardware is integrated into a cloud-based platform, exploiting the cloud computational powerto perform innovative algorithms for calibration (e.g., Machine Learning tools). Results and configurationscan be accessed through a web page without the installation of dedicated APPs or software. The electricalinput/output characteristic was measured with a dummy cell as a load, achieving excellent linearity.Furthermore, the device response to five different concentrations of potassium ferri/ferrocyanide redox probewas compared with a bench-top laboratory instrument. No difference in analytical sensitivity was found.Also, some examples of application-specific tests were set up to demonstrate the use in real-case scenarios.In addition, Support Vector Machine algorithm was applied to semi-quantitative analyses to classify theinput samples into four classes, achieving an average accuracy of 98.23%. Finally, COVID-19 related testsare presented and discussed (PDF) IoT and Biosensors: A Smart Portable Potentiostat With Advanced Cloud-Enabled Features. Available from: https://www.researchgate.net/publication/355214115_IoT_and_Biosensors_A_Smart_Portable_Potentiostat_With_Advanced_Cloud-Enabled_Features [accessed Oct 25 2021]

Rapid Quantification of SARS-Cov-2 Spike Protein Enhanced with a Machine Learning Technique Integrated in a Smart and Portable Immunosensor

An IoT-WiFi smart and portable electrochemical immunosensor for the quantification of SARS-CoV-2 spike protein integrated with machine learning features was developed. The immunoenzymatic sensor is based on the immobilization of monoclonal antibodies directed to SARS-CoV-2 S1 subunit on Screen-Printed Electrodes functionalized with gold nanoparticles, the analytical protocol involving a single-step sample incubation. Immunosensor performance was assessed by validation carried out in viral transfer medium, which is commonly used for de-sorption of nasopharyngeal swabs. Remarkable specificity of the response was demonstrated by testing H1N1 Hemagglutinin from swine-origin influenza A virus and Spike Protein S1 from Middle East respiratory syndrome coronavirus. Machine learning was successfully used for data processing and analysis: different support vector machine classifiers were evaluated proving that algorithms affect the classifier accuracy. The test accuracy of the best classification model in terms of true positive/true negative sample classification was 97.3%. In addition, ML algorithm can be easily integrated into the developed cloud-based portable Wi-Fi device. Finally, the immunosensor was successfully tested using a third generation replicating incompetent lentiviral vector pseudotyped with SARS-CoV-2 spike glycoprotein, thus proving the applicability of the immunosensor to whole virus detection

Understanding the role of imidazolium-based ionic liquids in the electrochemical CO2 reduction reaction

The development of efficient CO 2 capture and utilization technologies driven by renewable energy sources is mandatory to reduce the impact of climate change. Herein, seven imidazolium-based ionic liquids (ILs) with different anions and cations were tested as catholytes for the CO2 electrocatalytic reduction to CO over Ag electrode. Relevant activity and stability, but different selectivities for CO2 reduction or the side H 2 evolution were observed. Density functional theory results show that depending on the IL anions the CO 2 is captured or converted. Acetate anions (being strong Lewis bases) enhance CO2 capture and H2 evolution, while fluorinated anions (being weaker Lewis bases) favour the CO2 electroreduction. Differently from the hydrolytically unstable 1-butyl-3-methylimidazolium tetrafluoroborate, 1-Butyl-3-Methylimidazolium Triflate was the most promising IL, showing the highest Faradaic efficiency to CO (>95%), and up to 8 h of stable operation at high current rates (−20 mA & −60 mA), which opens the way for a prospective process scale-up

A film-forming graphene/diketopyrrolopyrrole covalent hybrid with far-red optical features: Evidence of photo-stability

A dianiline derivative of a symmetric donor-acceptor-donor diketopyrrolopyrrole-based dye is employed for the two-sided covalent functionalization of liquid exfoliated few layers graphene flakes, through a direct arylation reaction. The resulting nanohybrid features the properties of a polymeric species, being solution-processed into homogeneous thin films, featuring a pronounced red-shift of the main absorption band with respect to the model dye unit and energy levels comparable to those of common diketopyrrolopyrrole-based polymers. A good electrical conductivity and the absence of radical signals generated after intense white light illumination, as probed through electron paramagnetic resonance, suggest a possible future application of this composite material in the field of photoprotective, antistatic layers with tunable colors