Impedimetric Biosensor Research Articles

Label-free impedimetric biosensor based on a novel DNA-type receptor for selective determination of lactoferrin in human saliva

We propose a new impedimetric biosensor utilizing a unique H2N-(CH2)6-5’[TAGAGGATCAAA-AAA]4TAGAGGATCAAA3’ dsDNA oligonucleotide as a biorecognition element for selective lactoferrin determination in human saliva. The spatial orientation of the biosensing layer was defined by the immobilization of dsDNA oligonucleotide via H2N- group at 5’ end. The fabricated biosensors were characterized using both faradaic and non-faradaic models and proposing the Randles-based electrical equivalent circuits. Analysis of experimental data employing faradaic model equivalent circuits revealed a charge transfer resistance parameter R3 f dependent on the lactoferrin concentration. Hence, the biosensor response was established as the relationship between the ratio before/after bioreceptor-analyte interaction and the analyte concentration. The linear response of the developed impedimetric biosensors for laboratory samples was in the lactoferrin concentration range up to 625 nM with limit of detection of 1.25 nM. Notably, this study marked the pioneering application of these biosensors for quantitative lactoferrin detection in human saliva samples. The results exhibited accordance with the surface plasmon resonance method utilizing the same DNA-type receptor applied and colorimetric immunoassay. The presented impedimetric DNA-based biosensor for lactoferrin addresses the pressing need for rapid, simple, and effective analytical methods of immunomodulators detection with applicatory potential in clinical diagnosis.

A new immunosensing platform based on conjugated Poly(ThidEp-co-EDOT) copolymer for resistin detection, a new obesity biomarker.

Thedesign of a novel electrochemical impedimetric biosensor for label-free analysis of resistin, a biomarker for obesity, is reported. For the fabrication of the immunosensor, a novel approach composed of electrochemical copolymerization of double epoxy groups-substituted thiophene (ThidEp) and 3,4-Ethylenedioxythiophene (EDOT) monomers was utilized. Anti-resistin antibodies were covalently attached to the copolymer-coated electrode. The capture of resistin antigens by anti-resistin antibodies caused significant variations in charge transfer resistance (Rct) because of the immunoreactions between these proteins. Under optimum experimental variables, the changes in impedance signals were employed for the determination of resistin antigen concentration, and the prepared immunosensor based on conjugated copolymer illustrated a wide linear range between 0.0125 and 22.5 pg/mL, a low detection limit (LOD) of 3.71 fg/mL, and a good sensitivity of 1.22 kΩ pg-1mL cm2. The excellent analytical performance of the resistin immunosensor in terms of selectivity, sensitivity, repeatability, reproducibility, storage stability, and low detection limit might be attributed to the conductive copolymer film layer generation on the disposable indium tin oxide (ITO) platform. The capability of this system for the determination of resistin in human serum and saliva samples was also tested. The immunosensor results were in accordance with the enzyme-linked immunosorbent assay (ELISA) results. The matrix effects of human serum and saliva were also investigated, and the proposed immunosensor displayed good recovery ranging from 95.91 to 106.25%. Theengineered immunosensor could open new avenues for obesity monitoring.

An overview of electrochemical biosensors used for COVID-19 detection.

This short review presents the latest advances in the field of electrochemical biosensors, focusing particularly on impedimetric biosensors for the direct measurement of analytes. As a source of study we have chosen to describe these advances in the latest global health crisis originated from the COVID-19 pandemic, initiated by the SARS-CoV-2 virus. In this period, the necessity for swift and precise detection methods has grown rapidly due to an imminent need for the development of an analytical method to identify and isolate infected patients as an attempt to control the spreading of the disease. Traditional approaches such as the enzyme-linked immunosorbent assay (ELISA), were extensively used during the SARS-CoV-2 pandemic, but their drawbacks, including slow response time, became evident. In this context, the potential of electrochemical biosensors as an alternative for COVID-19 detection was emphasized. These biosensors merge electrochemical technology with bioreceptors, offering benefits such as rapidity, accuracy, portability, and real-time result provision. Additionally, we present instances of electrochemical biosensors modified with conductive polymers, eliminating the necessity for an electrochemical probe. The adaptability of the developed materials and devices facilitated the prompt production of electrochemical biosensors during the pandemic, creating opportunities for broader applications in infectious disease diagnosis.

Harnessing glycofluoroforms for impedimetric biosensing.

Glycans play a major role in biological cell-cell recognition and signal transduction but have found limited application in biosensors due to glycan/lectin promiscuity; multiple proteins are capable of binding to the same native glycan. Here, site-specific fluorination is used to introduce protein-glycan selectivity, and this is coupled with an electrochemical detection method to generate a novel biosensor platform. 3F-lacto-N-biose glycofluoroform is installed onto polymer tethers, which are subsequently immobilised onto gold screen printed electrodes, providing a non-fouling surface. The impedance biosensing platform is shown to selectively bind cancer-associated galectin-3 compared to control glycans and proteins. To improve the analytical capability, Bayesian statistical analysis was deployed in the equivalent circuit fitting of electrochemical impedance spectroscopy data. It is shown that Markov Chain Monte Carlo (MCMC) analysis is a helpful method for visualising experimental irreproducibility, and we apply this as a quality control step.

Synthesis and characterization of a nanostructure conductive copolymer based on polyaniline and polylactic acid as an effective substrate in proteins impedimetric biosensing.

Despite of all the developments in DNA microarray technology, there is not sufficient knowledge about protein abundance or their function in processes such as proteolysis, phosphorylation. Therefore, there is a significant need for direct detection and quantification of proteins, especially in processes such as proteomics, drug design and disease prediction. The present work introduce the new generation of polymeric substrate based on polyaniline and, polylactic acid, which it was used for impedimetric sensor in detection of proteins in particular for bovine serum albumin (BSA). In this copolymerization, the polylactic acid-block-polyaniline copolymer (PLA-b-PANI) was synthesized to attach polylactic acid and polyaniline using epichlorohydrin as a coupling agent. The structure of synthesized compounds in all steps, were confirmed by FT-IR and, 1H-NMR. The thermal properties and, morphology were analyzed by DSC, TGA, and, SEM. Also the electrochemical characteristics of fabricated PLA-b-PANI electrode were investigated by Electrochemical Impedance Spectroscopy (EIS) and Cyclic Voltammetry (CV). The results demonstrated that morphology of the PLA-b-PANI is sphere shape nanoparticles with dimension less than 100 nanometer diameters and, reasonable thermal properties. PLA-b-PANI was used to modify a screen-printed carbon electrode (SPCE) to fabricate a BSA impedimetric sensor. In order to increase the performance of the proposed impedimetric sensor, optimization of incubation time, pH and amount of PLA-b-PANI were investigated. The results show that the impedimetric sensor has the highest response when the electrode surface is covered with 5 microliters of PLA-b-PANI, and is incubated in BSA solution with pH 6.5 for 5 min. Impedimetric results showed that the PLA-b-PANI has excellent properties in reducing the charge transfer resistance and increasing the electron charge transfer rate. The final impedimetric sensor exhibited good repeatability, reproducibility, and chemical stability within the linear concentration range of 0.1-20 μg L-1 of BSA, and a detection limit of 0.05 μg L-1.

Electrochemical Paper-Based Nanobiosensor for Rapid and Sensitive Detection of Monkeypox Virus.

Monkeypox virus (MPXV) infection was classified as a public health emergency of international concern by the World Health Organization (WHO) in 2022, being transmitted between humans by large respiratory droplets, in contact with skin lesions, fomites, and sexually. Currently, there are no available accessible and simple-to-use diagnostic tests that accurately detect MPXV antigens for decentralized and frequent testing. Here, we report an electrochemical biosensor to detect MPXV antigens in saliva and plasma samples within 15 min using accessible materials. The electrochemical system was manufactured onto a paper substrate engraved by a CO2 laser machine, modified with gold nanostructures (AuNS) and a monoclonal antibody, enabling sensitive detection of A29 viral protein. The diagnostic test is based on the use of electrochemical impedance spectroscopy (EIS) and can be run by a miniaturized potentiostat connected to a smartphone. The impedimetric biosensing method presented excellent analytical parameters, enabling the detection of A29 glycoprotein in the concentration ranging from 1 × 10-14 to 1 × 10-7 g mL-1, with a limit of detection (LOD) of 3.0 × 10-16 g mL-1. Furthermore, it enabled the detection of MPXV antigens in the concentration ranging from 1 × 10-1 to 1 × 104 PFU mL-1, with an LOD of 7.8 × 10-3 PFU mL-1. Importantly, no cross-reactivity was observed when our device was tested in the presence of other poxvirus and nonpoxvirus strains, indicating the adequate selectivity of our nanobiosensor for MPXV detection. Collectively, the nanobiosensor presents high greenness metrics associated with the use of a reproducible and large-scale fabrication method, an accessible and sustainable paper substrate, and a low volume of sample (2.5 μL), which could facilitate frequent testing of MPXV at point-of-care (POC).

Green synthesized silver nanoparticles functionalized interdigitated electrodes for bacterial sensing using non-faradaic electrochemical impedance spectroscopy

In this study, we present a label-free non-faradaic impedimetric biosensor to detect bacterial cells using microfabricated gold interdigitated electrode (IDE). Silver nanoparticles (AgNP) are green synthesized using aqueous neem extract and characterized using Attenuated Total Reflectance- Fourier Transform Infrared spectra (ATR-FTIR), Dynamic Light Scattering (DLS), Scanning Electron Microscopy (SEM), and UV–Visible spectroscopy techniques. The synthesized AgNPs are well dispersed with an average size of 84 nm and showed an extensive antibacterial property indicated by a standard bioassay against Escherichia coli (E. coli). Gold IDEs are microfabricated by lithography on borosilicate glass wafers. The biofunctionalization of gold IDE is carried out using thiol‑gold covalent chemistry with mercaptohexanol (MCH). The self-assembled monolayer (SAM) of MCH facilitates drop-cast deposition of AgNP on the surface forming an MCH-AgNP. The functionalized IDE is electrochemically stable for further experiments and was validated by open circuit potential measurements. The objective of developing a label-free approach is confirmed by cyclic voltammetry analysis. Non-faradaic electrochemical impedance spectroscopy (nf-EIS) is carried out to detect E.coli cells suspended in water. The antibacterial property of AgNP is exploited to detect the decrease in cell concentration using nf-EIS. The impedance signatures corresponding to the trapping of cells are recorded with respect to time. Bacterial growth is a major challenge in maintaining water quality. The results demonstrated in this work would help to mitigate this problem effectively in a quick time without the need for skilled labor and sophisticated instruments required in traditional antibacterial testing.

Towards the rapid detection of haze‐forming proteins

Protein haze in white wine can be a serious quality defect because consumers perceive hazy wines as “spoiled”. Unfortunately, a specific method for the detection, or selective treatment, of such proteins in affected wines does not exist. Herein we investigate on the development of an easy‐to‐use sensor device that allows detection of haze-forming proteins (HFPs). Such a device is expected to overcome the limitations of the “heat test” currently used to assess the protein content in wine and the amount of bentonite needed to remove such proteins. To this aim, three different approaches were explored. Firstly, an impedimetric immunosensor against chitinases was developed and its performance assessed. Secondly, the exploitation of the dual role of HFPs as biorecognition element and analyte to develop an impedimetric biosensor was evaluated, in what can be considered a very unique strategy, representing a new paradigm in biosensing. Lastly, Fourier transform infrared (FT-IR) spectra were collected for various wine samples and chemometric tools such as discrete wavelet transform (DWT) and artificial neural networks (ANNs) were used to achieve the quantification of HFPs. Detection of HFPs at the μg/L level was achieved with both impedimetric biosensors, whereas the FT-IR-based approach allowed their quantification at the mg/L level in wine samples directly. The sensitivity of the developed methods may enable the rapid assessment of wine protein content.

Boron-doped diamond nanosheet volume-enriched screen-printed carbon electrodes: a platform for electroanalytical and impedimetric biosensor applications

This paper focuses on the development of a novel electrode based on boron-doped diamond nanosheet full-volume-enriched screen-printed carbon electrodes (BDDPE) for use as an impedimetric biosensor. Impedimetric biosensors offer high sensitivity and selectivity for virus detection, but their use as point-of-care devices is limited by the complexity of nanomaterials’ architecture and the receptor immobilisation procedures. The study presents a two-step modification process involving the electroreduction of diazonium salt at the BDDPE and the immobilisation of antibodies using zero-length cross-linkers for a selective impedimetric biosensor of Haemophilus influenzae (Hi). The incorporation of diamond nanosheets into BDDPE leads to enhanced charge transfer and electrochemical behaviour, demonstrating greatly improved electrochemically active surface area compared with unmodified screen-printed electrodes (by 44% and 10% on average for [Ru(NH3)6]Cl2 and K3[Fe(CN)6], respectively). The presented sensing system shows high specificity towards protein D in Hi bacteria, as confirmed by negative controls against potential interference from other pathogens, with an estimated tolerance limit for interference under 12%. The Hi limit of detection by electrochemical impedance spectroscopy was 1 CFU/mL (measured at − 0.13 V vs BDDPE pseudo-reference), which was achieved in under 10 min, including 5 min sample incubation in the presence of the analyte.Graphical abstract

Nanomaterials and Their Recent Applications in Impedimetric Biosensing.

Impedimetric biosensors measure changes in the electrical impedance due to a biochemical process, typically the binding of a biomolecule to a bioreceptor on the sensor surface. Nanomaterials can be employed to modify the biosensor's surface to increase the surface area available for biorecognition events, thereby improving the sensitivity and detection limits of the biosensor. Various nanomaterials, such as carbon nanotubes, carbon nanofibers, quantum dots, metal nanoparticles, and graphene oxide nanoparticles, have been investigated for impedimetric biosensors. These nanomaterials have yielded promising results in improving sensitivity, selectivity, and overall biosensor performance. Hence, they offer a wide range of possibilities for developing advanced biosensing platforms that can be employed in various fields, including healthcare, environmental monitoring, and food safety. This review focuses on the recent developments in nanoparticle-functionalized electrochemical-impedimetric biosensors.

Rapid Evaluation of Antibacterial Carbohydrates on a Microfluidic Chip Integrated with the Impedimetric Neoglycoprotein Biosensor.

The colonization of some bacteria to their host cell is mediated by selective adhesion between adhesin and glycan. The evaluation of antiadhesive carbohydrates in vitro has great significance in discovering new antibacterial drugs. In this paper, a microfluidic chip integrated with impedimetric neoglycoprotein biosensors was developed to evaluate the antibacterial effect of carbohydrates. Mannosylated bovine serum albumin (Man-BSA) was taken as the neoglycoprotein and immobilized on the microelectrode-modified gold nanoparticles (Au NPs) to form a bionic glycoprotein nanosensing surface (Man-BSA/Au NPs). Salmonella typhimurium (S. typhimurium) was selected as a bacteria model owing to its selective adhesion to the mannose. Electrochemical impedance spectroscopy (EIS) was used to characterize the adhesion capacity of S. typhimurium to the Man-BSA/Au NPs and evaluate the antiadhesive efficacy of nine different carbohydrates. It was illustrated that the 4-methoxyphenyl-α-D-pyran mannoside (Phenyl-Man) and mannan peptide (Mannatide) showed excellent antiadhesive efficacy, with IC50 values of 0.086 mM and 0.094 mM, respectively. The microfluidic device developed in this study can be tested in multiple channels. Compared with traditional methods for evaluating the antibacterial drug in vitro, it has the advantages of being fast, convenient, and cost-effective.

Enhanced Electrochemical Biosensing of the Sp17 Cancer Biomarker in Serum Samples via Engineered Two-Dimensional MoS2 Nanosheets on the Reduced Graphene Oxide Interface.

In the present investigation, we reported a label-free and highly effective immunosensor for the first time employing a nanostructured molybdenum disulfide nanosheets@reduced graphene oxide (nMoS2 NS@rGO) nanohybrid interface for the determination of sperm protein 17 (Sp17), an emerging cancer biomarker. We synthesized the nMoS2 NS@rGO nanohybrid using a one-step hydrothermal technique and then functionalized it with 3-aminopropyltriethoxysilane (APTES). Furthermore, the anti-Sp17 monoclonal antibodies were covalently attached to the APTES/nMoS2 NS@rGO/indium tin oxide (ITO) electrode utilizing 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-N-hydroxy succinimide (EDC-NHS) coupling chemistry. Bovine serum albumin (BSA) was then used to block nonspecific binding regions on the anti-Sp17/APTES/nMoS2 NS@rGO/ITO bioelectrode. The morphological and structural features of the synthesized nanohybrid and the modified electrodes were studied using transmission electron microscopy, scanning electron microscopy with energy dispersive X-ray (EDX) composition studies, atomic force microscopy, Fourier transform infrared spectroscopy, and Raman spectroscopy. The immunoreaction between the Sp17 antigen and anti-Sp17 antibodies on the surface of the BSA/anti-Sp17/APTES/nMoS2 NS@rGO/ITO sensing bioelectrode was applied as the basis for the detection technique, which measured the electrocatalytic current and impedimetric response change. The designed BSA/anti-Sp17/APTES/nMoS2 NS@rGO/ITO bioelectrode showed improved amperometric and impedimetric biosensing performance in the response studies, including remarkable sensitivity (23.2 μA ng-1mL cm-2 and 0.48 kΩ mL ng-1 cm-2), wider linearity (0.05-8 and 1-8 ng mL-1), an excellent lower detection limit (0.13 and 0.23 ng mL-1), and a rapid response time of 20 min. The biosensor exhibited impressive storage durability lasting 7 weeks and showed remarkable precision in identifying Sp17 in serum samples from cancer patients, as confirmed using the enzyme-linked immunosorbent assay method.

Part II: Impedance-based DNA biosensor for detection of isolated strains of phytopathogen Ralstonia solanacearum

In Part I, we demonstrated the complete development of a label-free, ultra-low sample volume requiring DNA-based biosensor to detect Ralstonia solanacearum, an aerobic non-spore-forming, Gram-negative, plant pathogenic bacterium, using non-faradaic electrochemical impedance spectroscopy (nf-EIS). We also presented the sensor's sensitivity, specificity, and electrochemical stability. In this article, we highlight the specificity study of the developed DNA-based impedimetric biosensor to detect various strains of R. solanacearum. We have collected seven isolates of R. solanacearum isolated from locally infected host plants (eggplant, potato, tomato, chilli, and ginger) from different parts of Goa, India. The pathogenicity of these isolates was tested on the eggplant, and the pathogen was confirmed by microbiological plating and polymerase chain reaction (PCR). We further report the insight into the DNA hybridization on the surface of Interdigitated Electrodes (IDEs) and the expansion of the Randles model for more accurate analysis. The interpretation of the sensor specificity is clearly demonstrated by the capacitance change observed at the electrode–electrolyte interface.

A Label-Free and Antibody-Free Molecularly Imprinted Polymer-Based Impedimetric Sensor for NSCLC-Cells-Derived Exosomes Detection.

In this study, a label-free and antibody-free impedimetric biosensor based on molecularly imprinting technology for exosomes derived from non-small-cell lung cancer (NSCLC) cells was established. Involved preparation parameters were systematically investigated. In this design, with template exosomes anchored on a glassy carbon electrode (GCE) by decorated cholesterol molecules, the subsequent electro-polymerization of APBA and elution procedure afforded a selective adsorption membrane for template A549 exosomes. The adsorption of exosomes caused a rise in the impedance of the sensor, so the concentration of template exosomes can be quantified by monitoring the impedance of GCEs. Each procedure in the establishment of the sensor was monitored with a corresponding method. Methodological verification showed great sensitivity and selectivity of this method with an LOD = 2.03 × 103 and an LOQ = 4.10 × 104 particles/mL. By introducing normal cells and other cancer cells derived exosomes as interference, high selectivity was proved. Accuracy and precision were measured, with an obtained average recovery ratio of 100.76% and a resulting RSD of 1.86%. Additionally, the sensors' performance was retained at 4 °C for a week or after undergoing elution and re-adsorption cycles seven times. In summary, the sensor is competitive for clinical translational application and improving the prognosis and survival for NSCLC patients.

Electrochemical aptasensors for pathogenic detection toward point-of-care diagnostics.

A biosensor system refers to a biomedical device, which detects biological, chemical, or biochemical components by converting those signals to an electrical signal by utilizing and uniting physical or chemical transducer with biorecognition elements. An electrochemical biosensor is generally based on the reaction of either production or consumption of electrons under a three-electrode system. Biosensor systems are exploited in a wide range of areas, such as medicine, agriculture, husbandry, food, industry, environment protection, quality control, waste disposal, and the military. Pathogenic infections are the third leading cause of death worldwide after cardiovascular diseases and cancer. Therefore, there is an urgent need for effective diagnostic tools to control food, water, and soil contamination result in protecting human life and health. Aptamers are peptide or oligonucleotide-based molecules that show very high affinity to their targets that are produced from large pools of random amino acid or oligonucleotide sequences. Generally, aptamers have been utilized for fundamental sciences and clinical implementations for their target-specific affinity and have been intensely exploited for different kinds of biosensor applications for approximately 30 years. The convergence of aptamers with biosensor systems enabled the construction of voltammetric, amperometric, and impedimetric biosensors for the detection of specific pathogens. In this review, electrochemical aptamer biosensors were evaluated by discussing the definition, types, and production techniques of aptamers, the advantages of aptamers as a biological recognition element against their alternatives, and a wide range of aptasensor examples from literature in the detection of specific pathogens.

COVID-19 impedimetric biosensor based on polypyrrole nanotubes, nickel hydroxide and VHH antibody fragment: specific, sensitive, and rapid viral detection in saliva samples

SARS-CoV-2 rapid spread required urgent, accurate, and prompt diagnosis to control the virus dissemination and pandemic management. Several sensors were developed using different biorecognition elements to obtain high specificity and sensitivity. However, the task to achieve these parameters in combination with fast detection, simplicity, and portability to identify the biorecognition element even in low concentration remains a challenge. Therefore, we developed an electrochemical biosensor based on polypyrrole nanotubes coupled via Ni(OH)2 ligation to an engineered antigen-binding fragment of heavy chain-only antibodies (VHH) termed Sb#15. Herein we report Sb#15-His6 expression, purification, and characterization of its interaction with the receptor-binding domain (RBD) of SARS-CoV-2 in addition to the construction and validation of a biosensor. The recombinant Sb#15 is correctly folded and interacts with the RBD with a dissociation constant (KD) of 27.1 ± 6.4 nmol/L. The biosensing platform was developed using polypyrrole nanotubes and Ni(OH)2, which can properly orientate the immobilization of Sb#15-His6 at the electrode surface through His-tag interaction for the sensitive SARS-CoV-2 antigen detection. The quantification limit was determined as 0.01 pg/mL using recombinant RBD, which was expressively lower than commercial monoclonal antibodies. In pre-characterized saliva, both Omicron and Delta SARS-CoV-2 were accurately detected only in positive samples, meeting all the requirements recommended by the World Health Organization for in vitro diagnostics. A low sample volume of saliva is needed to perform the detection, providing results within 15 min without further sample preparations. In summary, a new perspective allying recombinant VHHs with biosensor development and real sample detection was explored, addressing the need for accurate, rapid, and sensitive biosensors.

Development of disposable electrode for the detection of mosquito-borne viruses.

Development of disposable, rapid, and convenient biosensor with high sensitivity and reliability is the most desired method of viral disease prevention. To achieve this goal, in this work, a practical impedimetric biosensor has been implemented into a disposable electrode on a screen-printed carbon electrode (SPCE) for the detection of two mosquito-borne viruses. The biosensor fabrication has step-wisely carried out on the disposable electrode surface at room temperature: starting from conductive film formation, physical binding of the gold nanoparticles (AuNPs)-polyaniline (PAni) into the conductive film, and biofunctionalization. To get the maximum efficiency of the antibody, biotinylated antibody has been conjugated on the surface of AuNP-PAni/PAni-SPCE via the streptavidin-biotin conjugation method which is a critical factor for the high sensitivity. Using the antibody-antigen interaction, this disposable electrode has designed to detect mosquito-borne infectious viruses, Chikungunya virus (CHIKV), and Zika virus (ZIKV) separately in a wide linear range of 100fgmL-1 to 1ngmL-1 with a low detection limit of 1.33 and 12.31fgmL-1 , respectively.

A non-faradaic impedimetric biosensor for monitoring of caspase 9 in mammalian cell culture

Lower yields and poorer quality of biopharmaceutical products result from cell death in bioreactors. Such cell death is commonly associated with programmed cell death or apoptosis. During apoptosis, caspases are activated and cause a cascade of events that eventually lead to cell destruction. We report on an impedance spectroscopy measurement technique for the detection of total caspase-9 in buffer and complex fluids, such as cell culture media. Enhanced sensitivity is achieved by leveraging the physiochemical properties of zinc oxide and copper oxide at the electrode-solution interface. Characterisation of the biosensor surface was performed using scanning electron microscopy and indirectly using an enzyme-linked immunosorbent assay. The characteristic biomolecular interactions between the target analyte and specific capture probe of the biosensor are quantified using non-faradaic electrical impedance spectroscopy (nfEIS). The proof-of-concept biosensor demonstrated a detection limit of 0.07 U/mL (0.032 µM) in buffer. The sensor requires a low sample volume of 50 μL without the need for sample dilution facilitating rapid analysis. Using a luminescence-based assay, the presence of active caspase-9 was detected in the culture media following exposure to a pro-apoptotic agent. We envision that the caspase-9 biosensor will be useful as a cell stress screening device for apoptosis monitoring.

Graphene-Based Electrochemical Nano-Biosensors for Detection of SARS-CoV-2

COVID-19, a viral respiratory illness, is caused by Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2), which was first identified in Wuhan, China, in 2019 and rapidly spread worldwide. Testing and isolation were essential to control the virus’s transmission due to the severity of the disease. In this context, there is a global interest in the feasibility of employing nano-biosensors, especially those using graphene as a key material, for the real-time detection of the virus. The exceptional properties of graphene and the outstanding performance of nano-biosensors in identifying various viruses prompted a feasibility check on this technology. This paper focuses on the recent advances in using graphene-based electrochemical biosensors for sensing the SARS-CoV-2 virus. Specifically, it reviews various types of electrochemical biosensors, including amperometric, potentiometric, and impedimetric biosensors, and discusses the current challenges associated with biosensors for SARS-CoV-2 detection. The conclusion of this review discusses future directions in the field of electrochemical biosensors for SARS-CoV-2 detection, underscoring the importance of continued research and development in this domain.

Development of sensitive and stable electrochemical impedimetric biosensor based on T1R1 receptor and its application to detection of umami substances

Umami substances can provide a palatable flavour for food. In this study, an electrochemical impedimetric biosensor was developed for detecting umami substances. This biosensor was fabricated by immobilising T1R1 onto AuNPs/reduced graphene oxide/chitosan which was in advance electro-deposited onto a glassy carbon electrode. The evaluation by the electrochemical impedance spectrum method showed that the T1R1 biosensor performed well with low detection limits and wide linear ranges. Under the optimised incubation time (60 s), the electrochemical response was linearly related to the concentrations of the detected targets monosodium glutamate and inosine-5′-monophosphate within their respective linear range of 10-14 to 10-9 M and 10-16 to 10-13 M. The low detection limit of monosodium glutamate and inosine-5′-monophosphate was 10−15 M and 10−16 M, respectively. Moreover, the T1R1 biosensor exhibited high specificity to umami substances even in the real food sample. The developed biosensor still retained 89.24% signal intensity after 6-day storage, exhibiting a desirable storability.