Electrochemical Impedimetric Biosensor Research Articles

Ultrasensitive prostate cancer marker PCA3 detection with impedimetric biosensor based on specific label-free aptamers

Prostate cancer (PCa) appears among the most frequently diagnosed types of malignancies in males. Because of the high demand and increasing detection rate of early PCa, alongside the specificity limitations of the gold standard clinical tools available for the diagnosis and prognosis of prostate cancer, there is an urgent need for more reliable PCa markers and highly sensitive diagnostic tools to avoid under-treatment and over-diagnosis. PCA3, or prostate cancer antigen 3, is a potential prostate cancer biomarker that is more specific and useful for preventing unnecessary repeat biopsies, particularly in men with persistently high prostate-specific antigen indices after a negative biopsy. Additionally, an electrochemically based biosensor would prove to be a powerful diagnostic tool for PCA3 detection in urine because of its simplicity, sensitivity, and cost-effectiveness, in contrast to the more traditional PCa diagnostics that depend on blood testing. This paper aimed to design a novel and simple electrochemical impedimetric biosensor based on a label-free RNA-aptamer (CG3-PCA3) as the molecular recognition element for detecting PCA3. The proposed aptasensor for the detection of PCA3 has been developed using a screen-printed carbon electrode (SPCE) modified by gold nanoparticles (AuNPs), further improving sensitivity and allowing the immobilisation of thiolate aptamers on its surface. The findings presented here demonstrated a high sensitivity to PCA3, with a detection limit of 20 fM in artificial urine and 1 fM in buffer. These results indicate that the PCA3 aptasensor could be a promising tool for routine PCa diagnosis due to its high sensitivity and cost-effectiveness.

Impedimetric Detection of Cancer Markers Based on Nanofiber Copolymers.

The sensitive determination of folate receptors (FRs) in the early stages of cancer is of great significance for controlling the progression of cancerous cells. Many folic acid (FA)-based electrochemical biosensors have been utilized to detect FRs with promising performances, but most were complicated, non-reproducible, non-biocompatible, and time and cost consuming. Here, we developed an environmentally friendly and sensitive biosensor for FR detection. We proposed an electrochemical impedimetric biosensor formed by nanofibers (NFs) of bio-copolymers prepared by electrospinning. The biosensor combines the advantages of bio-friendly polymers, such as sodium alginate (SA) and polyethylene oxide (PEO) as an antifouling polymer, with FA as a biorecognition element. The NF nanocomposites were characterized using various techniques, including SEM, FTIR, zeta potential (ZP), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). We evaluated the performance of the NF biosensor using EIS and demonstrated FR detection in plasma with a limit of detection of 3 pM. Furthermore, the biosensor showed high selectivity, reliability, and good stability when stored for two months. This biosensor was constructed from 'green credentials' holding polymers that are highly needed in the new paradigm shift in the medical industry.

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.

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.

Recent Developments in Electrochemical-Impedimetric Biosensors for Virus Detection.

Viruses, including influenza viruses, MERS-CoV (Middle East respiratory syndrome coronavirus), SARS-CoV (severe acute respiratory syndrome coronavirus), HAV (Hepatitis A virus), HBV (Hepatitis B virus), HCV (Hepatitis C virus), HIV (human immunodeficiency virus), EBOV (Ebola virus), ZIKV (Zika virus), and most recently SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), are responsible for many diseases that result in hundreds of thousands of deaths yearly. The ongoing outbreak of the COVID-19 disease has raised a global concern and intensified research on the detection of viruses and virus-related diseases. Novel methods for the sensitive, rapid, and on-site detection of pathogens, such as the recent SARS-CoV-2, are critical for diagnosing and treating infectious diseases before they spread and affect human health worldwide. In this sense, electrochemical impedimetric biosensors could be applied for virus detection on a large scale. This review focuses on the recent developments in electrochemical-impedimetric biosensors for the detection of viruses.

The Use of Yeast Saccharomyces Cerevisiae as a Biorecognition element in the Development of a Model Impedimetric Biosensor for Caffeine Detection.

In the present study, an electrochemical-impedimetric biosensor using Saccharomyces cerevisiae as an effective biorecognition element was designed to detect caffeine. The presented biosensor consists of a previously developed stainless steel electrochemical cell constructed as a three-electrode system in the RCW side-by-side configuration. The electrochemical stability of the sensing electrode was evaluated by measuring the open circuit potential (OCP), and electrochemical impedance spectroscopy (EIS) was applied to determine the impedimetric response of the biosensor with Saccharomyces cerevisiae cells attached to the working electrode (WE) in the absence (0.9% NaCl) and presence (10 mg/mL in 0.9% NaCl) of caffeine. Moreover, the limit of detection (LOD) was determined. In this way, a new approach in biosensor development has been established, which involves assembling a low-cost and disposable electrochemical system to detect alkaloids such as caffeine. The developed biosensor represents a good candidate for detecting caffeine in beverages, foods, and drugs with the merits of time-saving, robustness, low cost, and low detection limit.

A label-free electrochemical impedimetric DNA biosensor for genetically modified soybean detection based on gold carbon dots.

A label-free electrochemical impedimetric biosensor was constructed based on gold carbon dots (GCDs) modified screen-printed carbon electrode for the detection of genetic modified (GM) soybean. The structure and property of GCDs were investigated. The GCDs can directly bind to single-stranded DNA probes through Au-thiol interaction and boost electric conductivity for the DNA sensor construction. The quantification of target DNA was monitored by the change of electron-transfer resistance (Ret) upon the DNA hybridization on sensor surface. Under the optimal conditions, the Ret response (vs. Ag reference electrode) increased with the logarithm of target DNA concentrations in a wide linear range of 1.0 × 10-7 - 1.0 × 10-13M with a detection limit of 3.1 × 10-14M (S/N = 3). It was also demonstrated that the proposed DNA sensor possessed high specificity for discriminating target DNA from mismatched sequences. Moreover, the developed biosensor was applied to detect SHZD32-1 in actual samples, and the results showed a good consistency with those obtained from the gel electrophoresis method. Compared with the previous reports for DNA detection, the label-free biosensor showed a comparatively simple platform due to elimination of complicated DNA labeling. Therefore, the proposed method showed great potential to be an alternative device for simple, sensitive, specific, and portable DNA sensor.

A novel, proof-of-concept electrochemical impedimetric biosensor based on extracellular matrix protein–adhesin interaction

Novel impedimetric biosensor for whole pathogen detection based on the extracellular matrix protein–adhesin interaction.

Electrochemical impedance biosensor for detection of saxitoxin in aqueous solution.

Saxitoxin is a cyanotoxin which is very harmful to human health; the concentration limit in drinking water is only 3μg/L. Therefore, a simple, fast, sensitive, low-cost, and specific method for its detection, quantification, and monitoring in water bodies is needed to avoid adverse effects on animal and human health. In this work, we developed an electrochemical impedimetric biosensor using a specific aptamer as recognition element for saxitoxin detection. This method allies the superior sensing characteristics of aptamers with the nondestructive, label-free, and easy working principles of the electrochemical impedance technique. The device presented sensitivity for detecting saxitoxin concentrations above 0.3μg/L, with high selectivity in negative control experiments, demonstrating a promising alternative for water toxin detection.

A label-free graphene-based impedimetric biosensor for real-time tracing of the cytokine storm in blood serum; suitable for screening COVID-19 patients.

Concurrent with the pandemic announcement of SARS-CoV-2 infection by the WHO, a variety of reports were published confirming the cytokine storm as the most mortal effect of the virus on the infected patients. Hence, cytokine storm as an evidenced consequence in most of the COVID-19 patients could offer a promising opportunity to use blood as a disease progression marker. Here, we have developed a rapid electrochemical impedance spectroscopy (EIS) sensor for quantifying the overall immune activity of the patients. Since during the cytokine storm many types of cytokines are elevated in the blood, there is no need for specific detection of a single type of cytokine and the collective behavior is just measured without any electrode functionalization. The sensor includes a monolayer graphene on a copper substrate as the working electrode (WE) which is able to distinguish between the early and severe stage of the infected patients. The charge transfer resistance (RCT) in the moderate and severe cases varies about 65% and 138% compared to the normal groups, respectively and a specificity of 77% and sensitivity of 100% based on ELISA results were achieved. The outcomes demonstrate a significant correlation between the total mass of the three main hypercytokinemia associated cytokines including IL-6, TNF-α and IFN-γ in patients and the RCT values. As an extra application, the biosensor's capability for diagnosis of COVID-19 patients was tested and a sensitivity of 92% and specificity of 50% were obtained compared to the RT-PCR results.

Electrochemical impedimetric biosensors, featuring the use of Room Temperature Ionic Liquids (RTILs): Special focus on non-faradaic sensing

Over the last decade, significant advancements have been made in the field of biosensing technology. With the rising demand for personalized healthcare and health management tools, electrochemical sensors are proving to be reliable solutions; specifically, impedimetric sensors are gaining considerable attention primarily due to their ability to perform label-free sensing. The novel approach of using Room Temperature Ionic Liquids (RTILs) to improve the sensitivity and stability of these detection systems makes long-term continuous sensing feasible towards a wide range of sensing applications, predominantly biosensing. Through this review, we aim to provide an update on current scientific progress in using impedimetric biosensing combined with RTILs for the development of sensitive biosensing platforms. This review also summarizes the latest trends in the field of biosensing and provides an update on the current challenges that remain unsolved.

An electrochemical impedimetric sensing platform based on a peptide aptamer identified by high-throughput molecular docking for sensitive l-arginine detection

As a primary building block for protein synthesis, l-arginine (l-Arg) is also a precursor for the synthesis of important metabolites, and is involved in various physiological and pathophysiological processes. l-Arg is a potential biomarker in clinical diagnosis and nutritional status assessment, making it valuable to quantify and monitor this biomolecule. In this study, peptide aptamers that specifically interact with l-Arg were identified by high-throughput molecular docking, and the binding capacities between the synthesized peptide aptamers and l-Arg were then measured by isothermal titration calorimetry. We hypothesized that the peptide aptamer with the greatest binding capacity could be used as the recognition element in a biosensor. A chemosynthetic peptide aptamer modified with mercaptan and spacer units (thioctic acid-GGGG-FGHIHEGY) was thus used to construct label-free electrochemical impedimetric biosensors for l-Arg based on gold electrodes. The optimum biosensor showed good sensitivity to l-Arg with a linear range of 0.1 pM–0.1 mM, and the calculated limit of detection (three times the signal-to-noise ratio) was 0.01 pM. Interference studies and assays of diluted serum samples were also carried out, and satisfactory results obtained. In conclusion, a potential method of peptide aptamer screening and biosensor fabrication for detecting small biological molecules was demonstrated.

Construction of an ultrasensitive electrochemical sensing platform for microRNA-21 based on interface impedance spectroscopy

A hybridization chain reaction (HCR) amplification-based electrochemical impedimetric biosensor is fabricated for the quick, sensitive, and specific detection of miRNA-21 (miR-21) via monitoring of electrode interfacial property changes in real-time. Two sequences of H1 and H2 are adopted to trigger HCR amplification. A large amount of linear DNA concatemer are formed which could change the interfacial properties of the electrode. Interfacial charge transfer resistance difference (Rct) is probed via electrochemical impedance spectroscopy (EIS) and Randles equivalent circuit. After amplifying via HCR, oligonucleotides with negatively charged repelling [Fe(CN)6]3−/4− ions can form a spatial blockage. HCR amplification strategy markedly enhanced the electrochemical signal with a limit of detection (LOD) down to 4.63 fM (S/N = 3). This strategy exhibited excellent selectivity for three different miRNAs: miR-199a, miR-141, and miR-155. Moreover, results show that the proposed method can be applied to miR-21 detection in the total RNA extracted from five cells. This work presents an enzyme-free and label-free EIS nucleic acid sensor for sensitively and selectively detecting miR-21, offering a promising approach in early diseases diagnosis.

Zirconium-porphyrin complex as novel nanocarrier for label-free impedimetric biosensing neuron-specific enolase

The development of impedimetric biosensors with ultrasensitive layers for the selective identification of cancer markers remains considerably challenging. Now, a novel electrochemical impedimetric biosensor based on the zirconium-porphyrin complex was constructed for the sensitive detection of neuron-specific enolase (NSE). Considering the efficient coordination of the Zr(III) and four N atoms bearing on porphyrin rings of 5,10,15,20-tetra(4-aminophenyl) porphyrin (TAPP) (denoted as Zr-TAPP), a Zr-TAPP complex was prepared by the solvothermal method. The Zr(III) coordination centers, amino groups, and π-π* bonds of the Zr-TAPP complex enabled strong affinity toward the antibody of NSE (AntiNSE). Compared with the conventional porphyrin-based Zr-MOF (PCN-224), the designed Zr-TAPP-based immunosensor displayed superior sensing performance for the NSE detection, thereby resulting in an extremely low detection limit of 7.1 fg mL−1 within a wide linear range of 10.0 fg mL-1-2.0 ng mL−1, accompanied with high selectivity and good stability. The prepared Zr-TAPP-based immunosensor can be further used to detect other cancer markers if anchoring other antibodies or probe molecules, thereby extending the application range of the metal-ligand complexes.

An impedimetric biosensor for E. coli O157:H7 based on the use ofself-assembled gold nanoparticles and protein G.

Two kinds of electrochemical impedimetric biosensors for the detection of E. coli O157:H7 are described and compared. They were fabricated using self-assembled layers of thiolated protein G (PrG-thiol) on (i) planar gold electrodes and (ii) gold nanoparticles (Au NPs) modified gold electrodes. The fabrications of the biosensors were characterized using cyclic voltammetry, electrochemical impedance spectroscopy, scanning electron microscopy and atomic force microscopy techniques. The modification of the planar gold electrode by Au NPs via self-assembled monolayer of 1,6-hexadithiol as a linker molecule increased the electrochemically active surface area by about 2.2 times. The concentration of PrG-thiol and its incubation time, as well as the concentration of IgG were optimized. The Au NP-based biosensor exhibited a limit of detection of 48 colony forming unit (cfumL-1) which is 3 times lower than that of the planar gold electrode biosensor (140cfumL-1). It also showed a wider dynamic range (up to 107cfumL-1) and sensitivity. The improved analytical performance of the Au NP-modified biosensor is ascribed to the synergistic effect between the Au NPs and the PrG-thiol scaffold. The biosensor exhibited high selectivity for E. coli O157:H7 over other bacteria such as Staphylococcus aureus and Salmonella typhimurium. Graphical abstract Schematic representations of sensor fabrication using Au NP-modified electrode (HKEC = heat- killed E. coli O157:H7).

Proximity binding-induced DNA assembly as signal translator and enzyme-catalyzed cleavage recycle as signal amplifier for highly sensitive electrochemical assay of target DNA

Based on target DNA (tDNA)-mediated proximity binding to develop unique DNA assembly as output translator and enzyme-cleaved recycle for signal amplification, an electrochemical impedimetric biosensor is developed. The DNA assembly is constructed by the proximal ligation of tDNA with two affinity probes (A1 and A2) on Au-Fe3O4 magnetic microspheres as substrate previously conjugated with A1, DNA linker (L1) and DNA transduction probe (TP). This binding brings the domain C2* of A2 close to the formed dsDNA TP:L1, facilitating the displacement of TP by C2* (C2*:L1). Under the T7-powered catalysis, the cleavage reaction, C2* liberation and next displacement of TP are initiated and successively recycled. In response to a single-target binding event, hundreds of output TP are released and are recognizable to hairpin DNA (NH2-HP) modified in the electrode surface. After exonuclease III (ExoIII) catalyzing the cleavage recycle, the retained C-rich repeats are converted into stable i-motif configuration when decreasing pH, generating decreased electron transfer impedance (Ret) owing to increased electrostatic attraction to [Fe(CN)6]3−/4−. Thus, the impedimetric biosensor is highly specific and sensitive with a limit of detection of 31 fM. This strategy would be promising to open up a new analytical route for disease diagnosis and bioanalysis.

Nanostructured titania based electrochemical impedimetric biosensor for non-invasive cancer detection

In the present work, the utilisation of bio-functionalised TiO2 nanoparticles has been reported for biosensing application especially in oral cancer detection. The bio-sensing electrode was fabricated on indium tin oxide coated conducting glass electrode using electrophoretic deposition technique. The proposed technique holds tremendous potential to make the approach effectively simple, label-free and most importantly a non-invasive technique. The prepared nanoparticles were characterised using x-ray diffractometer; Fourier transforms infrared spectroscopy, and x-ray photoelectron spectroscopy. In conjunction with this, the morphological studies of immunoelectrode (bovine serum albumin/anti-CYFRA-21-1/(3-aminopropyl)triethoxysilane/TiO2/indium tin oxide) were conducted by field emission scanning electron microscopy. The average roughness of various electrodes were investigated through atomic force microscopy. The biosensing properties of fabricated immunoelectrode were investigated using differential pulse voltammetry and cyclic voltammetry. However, the electrochemical response studies were carried out by using electrochemical impedance spectroscopy technique to measure the concentration of oral cancer biomarker (CYFRA 21-1). Additionally, we have also calculated the sensitivity and stability of fabricated immunoelectrode. It has been observed that the fabricated immunoelectrode shows a high sensitivity of 0.573 Ω mL ng−1, linear detection range of 0–12 ng mL−1, lower detection limit of 0.24 ng mL−1 and having a stability of 5 weeks.

Development of an Electrochemical Impedimetric Biosensor for the Detection of Telomerase Activity in Cancer Cells

In the last decade, a great amount of effort has gone into developing methods and devices that detect early signs of cancer diseases fast and with high efficiency. Telomerase based biosensors are an attractive alternative that comply with this requirements. Telomerase serves as a cancer biomarker because it is present in the vast majority of cancers. This enzyme elongates telomeres at the end of the chromosomal DNA, which causes cancer cells to become immortal. In this work, we developed a modified inter-digitated array gold microelectrode as the sensing platform with Electrochemical Impedance Spectroscopy (EIS) as the measuring technique for the telomerase activity. The resulting DNA biosensor detects telomerase activity in total protein extracts from Jurkat T-acute lymphoblastic leukemia cells. This simple system is a good proof of concept for a fast and efficient method that can detect telomerase activity in cancer cells.

Development of an Electrochemical Impedimetric Biosensor for the Detection of Telomerase Activity in Cancer Cells

Developing novel strategies to effectively detect biomarkers is of utmost importance in the early detection of cancer cells to increase the chances of a successful therapy. The proposed biosensor microchips works by tethering a biomolecular probe on the interdigital gold electrode array surfaces. This biomolecule probe can be customized according to the target biomarker, in order to recognize the change of conformation or bond upon contact. Telomerase is an enzyme responsible for the elongation of the telomeres between each cell division, restoring their length and avoiding cell death. This enzyme is responsible for the immortalization of cancer cells and it consists of a reverse transcriptase bound to a single stranded RNA (ssRNA) that is used as a template for telomere lengthening. The innovative research proposed in this project takes advantage of the telomerase and its ssRNA template by tethering a telomere-like ssDNA biomolecule probe that consists of a spacer composed of ten thymine, and the repeating sequence present in the telomeres of eukaryotic cells, 5’-TTAGGG-3’. The telomerase binds and elongates the probe, blocking electronic and capacitive processes on the microelectrode array surfaces. This change is detected by measuring the impedance using electrochemical impedance spectroscopy, which is correlated to the activity of telomerase and the presence of cancer cells. Moreover, telomerase is currently the target of many pharmaceutical companies to block cancer cell proliferation due to its presence in ca. 85% of known cancer cells (e.g. prostate, breast, and colorectal cancers). This project provides a unique opportunity to study the surface chemistry, impedance sensing, detection limits, and robustness of this microchip technology for its possible use at the point-of-care. This is funded by NSF Grant titled "Label-Free Electrochemical Capacitance DNA Sensing with Passive Wireless Radio Frequency Identification Sensor Technology" CHE-1152940.

An integrated dual functional recognition/amplification bio-label for the one-step impedimetric detection of Micro-RNA-21

Alteration in expression of miRNAs has been correlated with different cancer types, tumour stage and response to treatments. In this context, a structurally responsive oligonucleotide-based electrochemical impedimetric biosensor has been developed for the simple and sensitive detection of miRNA-21. A highly specific biotinylated DNA/LNA molecular beacon (MB) probe was conjugated with gold nanoparticles (AuNPs) to create an integrated, dual function bio-label (biotin-MB-AuNPs) for both biorecognition and signal generation. In the presence of target miRNA-21, hybridisation takes place resulting in the “activation” of the biotin-MB; this event makes the biotin group, which was previously “protected” by the steric hindrance of the MB stem-loop structure, accessible. The activated biotin-MB-AuNPs/miRNA complexes become available for capture, via supramolecular interaction, onto a nentravidin-modified electrode for electrochemical transduction. The binding event results in a decrease of the charge transfer resistance at the working electrode/electrolyte interface. The biosensor responded linearly in the range 1–1000 pM of miRNA-21, with a limit of detection of 0.3 pM, good reproducibility (Relative Standard deviation (RSD) =3.3%) and high selectivity over other miRNAs (i.e. miRNA-221 and miRNA-205) sequences. Detection of miRNA-21 in spiked serum samples at clinically relevant levels (low pM range) was also demonstrated, thus illustrating the potential of the biosensor for point-of-care clinical applications. The proposed biosensor design, based on the combination of a neutravidin transducing surface and the dual-function biotin-MB-AuNPs bio-label, provides a simple and robust approach for detection of short-length nucleic acid targets, such as miRNAs.