Surface Of Screen-printed Electrode Research Articles

Electrochemical Biosensor Enabling Sensitive Detection of miRNA Biomarkers

Biomarkers are becoming increasingly important for early diagnosis, exploration of responses to medication, and assessment of therapeutic efficacy of various diseases [1]. Increasing evidence shows that miRNAs can be employed as biomarkers for cancer and cardiovascular diseases. In this context, the development of powerful electrochemical diagnostic tools has grown in interest due to their sensitivity, speed, and low cost [1]. Additionally, electrochemical technologies offer a facile means to fabricate well-defined nanostructured surfaces in a controlled environment. The gold (Au)-based surface stands out for its ability to improve charge transfer characteristics, enhancing conductivity. It also serves as a stable substrate for further modification, enabling the detection of target analytes [2]. Here, we utilized an electrochemical platform by using a 3-electrode carbon-based screen-printed electrode (SPE) electrochemically modified with gold and subsequently attached a specific DNA-based redox probe to detect low levels of miRNAs as a biomarker. Our design was applied to detect miRNA-1 (5′-TGG AAT GTA AAG AAG TAT GTA T-3′), demonstrating exceptional sensitivity and a low detection limit (LOD) of 120 fM. We also discuss how the developed platform broadly benefits numerous other miRNA sequences. Device preparation method: A screen-printed electrode (SPE) consisting of a 3 mm diameter carbon working electrode, an Ag reference electrode, and a carbon counter electrode was used as a base for device development. The bare SPE was activated during 10 cycles of cyclic voltammetry (CV) with potentials ranging from -0.5 to +1 V at a scan rate of 20 mVs-1 using PBS as the electrolyte. Before gold electrodeposition, the activated SPE was rinsed with PBS and deionized water. The electrode was submerged in a solution of 115 mM HAuCl4 (in 0.5 M H2SO4) during 10 CV cycles with the same parameters described above. Next, the DNA-probe sequence was attached to the working surface of the Au-decorated screen-printed electrode (Au-SPE). The DNA-probe sequence was modified in advance with C6-S-S-R at the 5′ end (enabling covalent binding onto Au) and methylene blue (MB) at the 3′ end (allowing electron transport at the electrode). Before modification of SPE, the disulfide bond was reduced using tris(2-carboxyethyl)-phosphine hydrochloride) (TCEP) to enable covalently attachment of the HS-DNA-MB probe to the Au-SPE [2]. The attachment of the DNA-probe sequence was achieved by placing 10 µL of the diluted probe onto the working electrode area and allowing for 1 hour of modification at room temperature to take place, as shown in Fig. 1(A). Results & Discussion: The device was evaluated for analytical performance in the presence of increasing target concentrations, as shown in Fig. 1 (A). In the absence of a miRNA-1 target, the stem-loop structure of the DNA probe remains closed, forcing the redox MB-reporter into proximity to the electrode, allowing for efficient electron transport. In the presence of miRNA-1, the target molecule binds to a complementary sequence of the DNA probe, opening the loop. The structure assumes a relatively rigid conformation where MB is farther from the gold surface, reducing the observed signal, Fig. 1 (B). The observed current gradually decreases upon the successive addition of miRNA target molecules for both cathodic, Ipc, and anodic, Iac, peaks. Using the dependence of anodic peak, Iac, the sensor was calibrated as a function of added miRNA-1 concentration in the range between 0 and 4 nM. Using the calibration curve shown in Fig. 1 (C), the detection limit was estimated to be 120 femtoM. The device was further tested for stability and exhibited robust performance over an extended period displaying minimal fluctuations in the recorded current. While we demonstrate the sensitive detection of miRNA-1, the design can be easily adapted for other miRNA sequences [3]. Additionally, it introduces exciting prospects for multiplexing, where multiple miRNA sequences can be easily detected with only minor adjustments to the established design. Fig. 1 (A) Schematics of the device fabrication process and mechanism for the development of Smart Electrochemical Sensors setup; (B) Cyclic Voltammetry (CV) responses during Au electrodeposition (black) in the absence of miRNA-1 target (red) and presence of miRNA-1 target (blue); (C) The device response curve concerning successive addition of miRNA-1 target molecule into the device chamber (different concentrations of target from 0 to ~ 4000 pM), the inset shows CV response used to plot the calibration curve. Acknowledgment: NSF DMR 2204027, US DOE EPSCoR DE-SC0024284

Label free and ultrasensitive electrochemical detection of carcinoembryonic antigen using a disposable aptasensor based on Pd and graphene

We prepared an electrochemical aptasensor for label-free analysis of carcinoembryonic antigen (CEA). Herein, PdNPs were synthesized on the surface of graphene-modified screen-printed carbon electrodes (Pd-GN/SPCE) via electrochemical method for immobilisation of CEA aptamer as a bio-recognition element. The morphological characterizations of the fabricated Pd-GN/SPCE were performed using SEM, EDX, and elemental mapping that confirmed the successful electrochemical deposition of PdNPs. A disposable aptasensor for ultrasensitive electrochemical measurement of CEA was proposed by immobilising CEA aptamer on Pd-GN/SPCE. The electrochemical measurements were performed using 5 mM [Fe(CN)6]3-/4- as a redox probe. The electrochemical deposition of PdNPs improved the electrical conductivity resulting in enhanced current response for the redox probe. When CEA antigen interacts with CEA aptamer, it reduces the oxidation and reduction current of [Fe(CN)6]3-/4- due to the attachment of biomolecules to the electrode surfaces that hinders the electron transfer. The current changes of the redox species before and after CEA binding were measured using DPV. The developed aptasensor possessed a wide calibration range of 0.002–200 ng/mL and detection limit of 1.0 pg/mL. The aptamer-immobilised electrode provides an exceptional selectivity, reproducibility, and sensitive measurement of CEA. Moreover, the effective sensing of CEA in human serum samples with great recoveries (99.4––106 %) demonstrated its capability for clinical analysis.

Simultaneous speciation of inorganic arsenic (III and V) utilizing gold-manganese oxide nanoparticles modified electrochemical sensors

Inorganic arsenic is primarily present in two stable valence states: III and V, each exhibiting different toxicological profiles. Accurately distinguishing As(III) and As(V) is, therefore, crucial for precise risk assessment and environmental monitoring. The prevalent approach for such speciation commonly involves a combination of bulk and expensive high-performance liquid chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP-MS) instruments. In response to the demand for more streamlined and efficient techniques, several alternative methods, including voltammetry, have been explored. Despite the application of gold and manganese in the voltammetric detection of inorganic arsenic, the simultaneous measurements of As(III) and As(V) have not been solved. This study introduced a novel nanocomposite with an Au-Mn ratio of 10:1 (m/m) for modifying the surface of screen-printed carbon electrodes (SPCEs). These modified SPCEs were successfully applied in the monitoring and speciation of inorganic arsenic simultaneously by voltammetry during simulated wastewater treatment for the first time. The linear range is 0.2 to 2 mg·mL-1 (ppm) with the limits of detection (LOD) of 0.048 ppm for total arsenic and 0.07 ppm for As(III), respectively. Notably, the performance and accuracies of developed voltammetric sensors for simultaneous detection of total arsenic, As(III) and As(V) were confirmed through validation with the sophisticated HPLC coupled to triple quadrupole ICP-MS (HPLC-ICP-QQQ) method.

Electrochemical Immunoassay Platform for Human Monkeypox Virus Detection.

In this study, we reported a selective impedimetric biosensor for the detection of A29 which is the target protein of the monkeypox virus (MPXV). The working principle of the biosensor relies on the interaction mechanism between A29, which is an internal membrane protein of MPXV, and the heparan sulfate receptor. For this purpose, after immobilizing heparan sulfate onto the gold screen-printed electrode surface, its interaction with A29 protein was monitored using electrochemical impedance spectroscopy. After the optimization of experimental parameters, the analytical characteristics of the developed MPVX immunosensor were examined. The developed immunosensor exhibited a linear detection range between 2.0 and 50 ng mL-1, with a detection limit of 2.08 ng mL-1 and a quantification limit of 6.28 ng mL-1. Furthermore, a relative standard deviation value of 2.82% was determined for 25 ng mL-1. Apart from that, sample application studies were also performed with the standard addition of A29 protein to 1:10 diluted real serum samples that were taken from healthy individuals, and very good recovery values were obtained.

Functionalization of Carbon Electrode Surface Using Polyaniline and Gold Nanoparticles for Protein Immobilization

This study presents a screen-printed carbon electrode surface functionalization to immobilize proteins. The process involves the deposition of gold nanoparticles on the carbon electrode and subsequent surface modification to immobilize bovine serum albumin–fluorescein isothiocyanate conjugate (BSA-FITC). A combination of conducting polymer and gold nanoparticles was employed to enhance the electrode contact area and current transmission. The electrode surface morphology and properties were evaluated by scanning electron microscopy (SEM) and Raman spectroscopy, showing the porous polyaniline (PANI) film has been formed on the carbon electrode surface with the network of dendritic nanofibers. Gold nanoparticles were evenly distributed on the PANI film to form a nanocomposite material layer on the carbon electrode. The observed size of these gold nanoparticles was significantly smaller than those on the electrode without a PANI film. Additionally, the current transmission of this composite material layer has been verified by cyclic voltammetry (CV). The results show that the amplitude of the redox peak was doubled compared to the bare carbon electrode and gold nanoparticle-only deposited electrode. Moreover, the success of the process was verified by fluorescence and cyclic voltammetry. The fluorescent green color observed under the microscope indicated that BSA-FITC was on the electrode surface. The change of CV signal after each step verified the formation of a new layer on the electrode surface. These results are anticipated to play an important role in developing high-sensitivity immunosensors in the future.

Development of the BAT-26 mutation-based electrochemical genosensor for identifying microsatellite instability in relationship to cancer

Screening for microsatellite instability is one of the biggest challenges in the cancer diagnosis due to its association with different types of cancer development. In this sense, the development of fast and highly sensitive analytical devices for early the cancer detection is a high priority. In the present work, a highly sensitive electrochemistry genosensor is developed and described for BAT-26 mutation detection using: differential pulse voltammetry (DPV), and cyclic voltametric (CV) supported by scanning tunnel microscopy (STM). The redox reaction of Fe(CN)63−/4– was monitored to probe the presence DNA target. The results demonstrate that modifying the screen-printed gold electrode surface (SPGE) 250AT with a combination of 11-mercaptoundecanoic acid (MUA) and a single-stranded DNA oligonucleotide successfully altered the electrode's properties, particularly its affinity for detecting the BAT-26 mutation under physiological conditions. The genosensor exhibited a wide linear range (100 aM to 1 μM) and achieved a low detection limit of 71.74 aM under optimal parameters. These characteristics, coupled with its high sensitivity and rapid analysis time, make it a promising candidate for development into a portable microsatellite instability diagnostic kit.

Label-free aptasensor targeting Staphylococcus aureus surface Protein A

The rapid detection of Staphylococcus aureus (S. aureus) is of major importance for the biomedical world in the context where the antimicrobial resistance (AMR), the “silent pandemic”, is considered a global public health threat to humanity. There are various approaches to detect S. aureus either as a whole bacterium or via different biomarkers that indicate the presence of this microorganism. Herein, a label-free aptasensor was designed for the detection of a cellular bacterial wall component, protein A (PrA), as a biomarker for the bacteria presence. A rapid procedure, under 5 min, that involves the deposition of the aptamer at the surface of a gold-based screen-printed electrode was applied followed by a blocking step to prevent non-specific adsorption of the target. After the incubation with PrA, the electrochemical detection was achieved, the aptasensor exhibiting good sensitivity across a wide linear range from 25 nM to 1000 nM, with a limit of detection (LOD) of 8.33 nM in standard solutions of the target. S. aureus was detected in commercial human serum and culture samples displaying a good correlation of the signal and the number of bacteria colonies, in the latter case.

Point-of-care dengue detection: polydopamine-modified electrode for rapid NS1 protein testing for clinical samples.

Early diagnosis of dengue infection by detecting thedengue virus non-structural protein 1 (DENV-NS1) is important to the patients to initiate speedy treatment. Enzyme-linked immunosorbent assay (ELISA)-based NS1 detection and RT-PCR are time-consuming and too complex to be employed in remote areas of dengue-endemic countries. Meanwhile, those of NS1 rapid test by lateral flow assay suffer from low detection limit. Electrochemical-based biosensors using screen-printed gold electrodes (SPGEs) have become a reliable detection method to convey both ELISA's high sensitivity and rapid test portability. In this research, we developed an electrochemical biosensor for DENV-NS1 detection by employing polydopamine (PDA)-modified SPGE. The electrodeposition of PDA on the surface of SPGE serves as a bioconjugation avenue for anti-NS1 antibody through a simple and low-cost immobilization procedure. The biosensor performance was evaluated to detect DENV-NS1 protein in PBS and human serum through a differential pulse voltammetric (DPV) technique. The developed sensing platform displayed a low limit of detection (LOD) of 1.63pgmL-1 and a wide linear range of 10pgmL-1 to 1ngmL-1 (R2 ∼ 0.969). The sensing platform also detected DEV-NS1 from four different serotypes in the clinical samples collected from dengue patients in India and Indonesia, with acceptable sensitivity, specificity, and accuracy values of 90.00%, 80.95%, and 87.65%, respectively. This result showcased the facile and versatile method of PDA coating onto the surface of screen-printed gold electrodes for a miniaturized point-of-care (PoC) detection device.

Reduced Graphene Oxide Modified Enzyme Inhibition-Based Biosensor System for Detection of Paraoxon as a Nerve Agent Simulant

Nerve agents, including organophosphorus compounds such as paraoxon, are potent and highly toxic chemicals with grave implications for human health and the environment. In this paper, we present the development of a novel enzyme inhibition-based biosensor for the sensitive and selective detection of paraoxon, which is commonly used as a surrogate for nerve agents. The biosensor employs reduced graphene oxide as a screen-printed electrode surface modification nanomaterial, leading to increased surface electroactivity and, thus, more sensitive detection. The biosensor exhibits a low detection limit of 0.56 pg/ml (limit of detection, LOD) and 1.25 pg/ml (limit of quantification, LOQ), highlighting its high sensitivity for trace-level analysis of nerve agents in complex sample matrices. Our biosensor demonstrates remarkable selectivity for paraoxon, with minimal interference from other non-target chemicals. Stability and repeatability tests reveal that the system maintains its performance integrity over a 45-day period and consistently produces readings with a margin of error of only 5%. Real sample testing in river water, wastewater, and tap water further confirms the biosensor's practical utility, with recovery percentages ranging from 84 to 115%. This biosensor represents a significant advancement in biosensor technology, facilitating the rapid, cost-effective, and reliable detection of toxic substances in real-world scenarios.

Electrochemically assisted DNA and thioaromatic assembly as sensing and antifouling interface for food allergens.

Food allergies have become a global issue and are estimated to affect approximately 220 million people worldwide. Allergy to peanuts can easily become life-threatening and induce anaphylactic reactions. Mislabeling and cross-contamination during food processing can occur in the frame of world population growth and pose a serious health issue. As the mandatory allergen list is not uniform worldwide, the development of routine analytical strategies with high specificity and sensitivity is a demanding task to aid in the rapid identification of allergenic foods. In this work, an electrochemical aptasensor for Ara h1 peanut allergen was developed by immobilizing the specific aptamer by the inserting method. First, a layer of p-aminothiophenol (p-ATP) was immobilized on the gold surface of screen-printed electrodes (GSPE) to improve the aptamer insertion and reduce the fouling effects at the electrode surface. The grafting of the p-ATP and Ara h1 aptamer on the GSPE surface was monitored by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). The resulting disposable aptasensor allowed for indirect electrochemical detection of Ara h1 protein in the presence of 5 mM ferro/ferricyanide as a redox probe. The electrochemical response upon aptamer-target interaction was monitored in the concentration range 1-250 nM, and two limits of detection in the nanomolar range were estimated based on DPV (2.78 nM Ara h1) and EIS (0.82 nM Ara h1) measurements. The aptasensor was successfully applied to real sample analysis.

An applied potential powered screen-printed electrode for sensitive EC-SERS detection of acetamiprid

Acetamiprid (AAP) is a common neonicotinoid pesticide and their residues have emerged as a widely concerned issue on vegetables and fruits. In this work, a facile electrochemical surface-enhanced Raman spectroscopy (EC-SERS) method with one step of “mixing and detection” was proposed for detection of AAP. Wherein, screen printed electrode (SPE) was modified by dropping silver nanoparticles (AgNPs) onto the SPE surface. Compared to the normal SERS detection, the EC-SERS had a 5-fold enhancement in SERS signal with an applied potential at –0.7 V. Under the optimal applied potential, a wide linear range from 0.1 μM to 1000 μM was achieved with a low limit of detection (LOD, 0.04 μM), about 7-fold lower than that of the normal SERS method (LOD, 0.31 μM). Density functional theory (DFT) calculation revealed that the van der Waals force between AAP molecules and AgNPs was greatly enhanced. Besides, the EC-SERS method was successfully applied for the detection of AAP in Brassica chinensis L., and the results correlated well (R2 =0.9760) with those obtained from liquid chromatography-tandem mass spectrometry (HPLC-MS). The proposed method featured in easy operation, rapid detection and high sensitivity, which can act as a promising tool for monitoring pesticide residues in food samples.

An electrochemically synthesized molecularly imprinted polymer for highly selective detection of breast cancer biomarker CA 15-3: a promising point-of-care biosensor.

In this study, a molecularly imprinted polymer film (MIP) was prepared on the surface of a disposable carbon screen-printed electrode (C-SPE) using (3-acrylamidopropyl)trimethylammonium chloride (AMPTMA) as a functional monomer and the cancer biomarker carbohydrate antigen 15-3 (CA 15-3) as a template. The MIP was synthesized by in situ electropolymerization (ELP) of the AMPTMA monomer in the presence of the CA 15-3 protein on the C-SPE surface. The target was subsequently removed from the polymer matrix by the action of proteinase K, resulting in imprinted cavities with a high affinity for CA 15-3. Electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterize the different phases of the sensor assembly. Chemical and morphological analysis was performed using RAMAN and scanning electron microscopy (SEM). CA 15-3 was successfully detected in a wide working range from 0.001 U mL-1 to 100 U mL-1 with a correlation coefficient (R2) of 0.994 in 20 min. The MIP sensor showed minimal interference with other cancer proteins (CEA and CA 125). Overall, the developed device provides a rapid, sensitive, and cost-effective response in the detection of CA 15-3. Importantly, this comprehensive approach appears suitable for point-of-care (PoC) use, particularly in a clinical context.

A TiO2-adenine nanocomposite as modification material for screen-printed gold electrode to detect H1N1 (swine flu) virus: A disposable genosensor

A genosensor based on TiO2-adenine nanocomposites (NCs) modified screen-printed gold electrode surface (TA-SPGE) was immobilized with haemagglutinin (HA) gene specific probe containing 5′ amine group and investigated for swine flu virus (H1N1). Bare SPGE surface was modified by drop coating of the nanocomposites. The NCs and precursors were characterized structurally and morphologically by adopting various techniques such as Fourier Transform Infrared (FTIR) spectra, X-ray Diffraction (XRD) spectra, UV–Visible spectra, Raman spectra, Zeta sizer, Field-Emission Scanning Electron Microscopy (FE-SEM), Energy Dispersive spectra (EDS) and High-Resolution Transmission Electron Microscopy (HR-TEM) with Selected Area Electron Diffraction (SAED) pattern. Electrochemical techniques termed as cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectra (EIS) were employed for analytical detection of single-stranded viral genome in phosphate buffer saline (pH-7.2) by using methylene blue (MB) redox indicator. Moreover, the performance of designed electrode was optimized at different concentration, pH, and scan rates. The proposed genosensor revealed high sensitivity 10.6 μA/ng.cm2, lower detection limit (LOD) of 6.7 × 10-8 ng/mL and wide linear range of 3 × 10-8-3 × 10-3 ng/mL with regression coefficient of R2 = 0.989 towards swine flu virus DNA. Additionally, the modified genosensor revealed good stability and specificity against human DNA and various pathogens such as H3N2 influenza virus, Bacillus subtilis, Neisseria meningitidis, Staphylococcus aureus and Streptococcus pyogenes. The proposed genosensor is based on relatively simple methodology, quite amicable and could promptly be used as point-of-care test for swine flu virus detection.

Effect of Aspect Ratio of a Gold-Nanorod-Modified Screen-Printed Carbon Electrode for Carbaryl Detection in Three Different Samples of Vegetables.

In this study, three different sizes of gold nanorods (AuNRs) were synthesized using the seed-growth method by adding various volumes of AgNO3 as 400, 600, and 800 μL into the growth solution of gold nanoparticles. Three different sizes of AuNRs were then characterized using UV-vis spectroscopy, high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED) patterns, and atomic force microscopy (AFM) to investigate the surface morphology, topography, and aspect ratios of each synthesized AuNR. The aspect ratios from the histogram of size distributions of three AuNRs as 2.21, 2.53, and 2.85 can be calculated corresponding to the addition of AgNO3 volumes of 400, 600, and 800 μL. Moreover, each AuNR in three different aspect ratios was drop-cast onto the surface of a commercial screen-printed carbon electrode (SPCE) to obtain three different SPCE-modified AuNRs (SPCE-A400, SPCE-A600, and SPCE-A800, respectively). All SPCE-modified AuNRs were then evaluated for their electrochemical behavior using cyclic voltammetry and electrochemical impedance spectroscopy (EIS) techniques and the highest electrochemical performance was shown as the order of magnitude of SPCE-A400 > SPCE-A600/SPCE-A800. The reason for the highest electrocatalytic activity of SPCE-A400 might be due to the smallest particle size and uniform distribution of AuNRs ∼ 2.2, which enhanced the charge transfer, thus providing the highest electroactive surface area (0.6685 cm2) compared to other electrodes. These results also confirm that the sensing mechanism for all SPCE-modified AuNRs is controlled by diffusion phenomena. In addition, the optimum pH was obtained as 4 for carbaryl detection for all SPCE-modified AuNRs with the highest current shown by SPCE-A400. Furthermore, SPCE-A400 has the highest fundamental parameters (surface coverage, catalytic rate constant, electron transfer rate constant, and adsorption capacity) for carbaryl detection, which were investigated using cyclic voltammetry and chronoamperometric techniques. The electroanalytical performances of all SPCE-modified AuNRs for carbaryl detection were also investigated with SPCE-A400 displaying the best performance among other electrodes in terms of its linearity (0.2-100 μM), limit of detection (LOD) ∼ 0.07 μM, and limit of quantification (LOQ) ∼ 0.2 μM. All SPCE-modified AuNRs were also subsequently evaluated for their stability, reproducibility, and selectivity in the presence of interfering species such as NaNO2, NH4NO3, Zn(CH3CO2)2, FeSO4, diazinon, and glucose and show reliable results as depicted from %RSD values less than 3%. At last, all SPCE-modified AuNRs have been employed for carbaryl detection using a standard addition technique in three different samples of vegetables (cabbage, cucumber, and Chinese cabbage) with its results (%recovery ≈ 100%) within the acceptable analytical range. In conclusion, this work demonstrates the great potential of a disposable device based on an AuNR-modified SPCE for rapid detection and high sensitivity in monitoring the concentration of carbaryl as a residual pesticide in vegetable samples.

Redox probe-free electrochemical immunosensor utilizing electropolymerized melamine on reduced graphene oxide for the point-of-care diagnosis of gastric cancer

Pepsinogen I (PG I) is a biomarker that plays a crucial role in the diagnosis of gastric cancer. The development of biosensor to monitor PG I overexpression in serum is crucial for early gastric cancer diagnosis, offering a less invasive alternative to the costly and uncomfortable gastroscopy procedure. This study presents a cost-efficient, scalable and disposable label-free biosensing strategy for detecting PG I, utilizing a redox-active polymelamine electrodeposited on a reduced graphene oxide screen-printed electrode surface (PM-rGO/SPE). Under optimized conditions, the conducting polymer PM was deposited on the rGO/SPE via a potentiodynamic method. The structural and morphological features of PM-rGO/SPE were analyzed with the assistance of Raman and Scanning Electron Microscopy analysis. Specific monoclonal anti-PG I antibodies were immobilized on the in situ prepared redox-active layer via EDC/NHS chemistry to develop a novel electrochemical immunosensor. Unlike the traditional immunosensing strategies which utilizes external redox probe solution for measuring the signal, the developed configuration allowed for redox-probe free monitoring of current changes of the redox active PM resulting from the formation of the immunocomplex on the electrode surface. Utilizing this method, PG I detection spanned a clinically relevant concentration range of 0.01–200 ng/mL, with a low limit of detection at 9.1 pg/mL. The electrochemical immunosensor demonstrated specificity against other biomarkers such as PDCD1, ErBb2, and CD28 with negligible interference. The immunosensor exhibited excellent recovery capabilities for PG I detection in serum samples. These findings underscore the potential of the PM-rGO/SPE immunosensor as a point-of-care diagnostic tool for gastric cancer.

Carbon Nanostructured Immunosensing of Anti-SARS-CoV-2 S-Protein Antibodies.

The rampant spread and death rate of the recent coronavirus pandemic related to the SARS-CoV-2 respiratory virus have underscored the critical need for affordable, portable virus diagnostics, particularly in resource-limited settings. Moreover, efficient and timely monitoring of vaccine efficacy is needed to prevent future widespread infections. Electrochemical immunosensing poses an effective alternative to conventional molecular spectroscopic approaches, offering rapid, cost-effective, sensitive, and portable electroanalysis of disease biomarkers and antibodies; however, efforts to improve binding efficiency and sensitivity are still being investigated. Graphene quantum dots (GQDs) in particular have shown promise in improving device sensitivity. This study reports the development of a GQD-functionalized point-of-contamination device leveraging the selective interactions between SARS-CoV-2-specific Spike (S) Protein receptor binding domain (RBD) antigens and IgG anti-SARS-CoV-2-specific S-protein antibodies at screen-printed carbon electrode (SPCE) surfaces. The immunocomplexes formed at the GQD surfaces result in the interruption of the redox reactions that take place in the presence of a redox probe, decreasing the current response. Increased active surface area, conductivity, and binding via EDC/NHS chemistry were achieved due to the nanomaterial inclusion, with 5 nm, blue luminescent GQDs offering the best results. GQD concentration, EDC/NHS ratio, and RBD S-protein incubation time and concentration were optimized for the biosensor, and inter- and intra-screen-printed carbon electrode detection was investigated by calibration studies on multiple and single electrodes. The single electrode used for the entire calibration provided the best results. The label-free immunosensor was able to selectively detect anti-SARS-CoV-2 IgG antibodies between 0.5 and 100 ng/mL in the presence of IgM and other coronavirus antibodies with an excellent regression of 0.9599. A LOD of 2.028 ng/mL was found, offering comparable findings to the literature-reported values. The detection sensitivity of the sensor is further compared to non-specific IgM antibodies. The developed GQD immunosensor was compared to other low-oxygen content carbon nanomaterials, namely (i) carbon quantum dot (CQD), (ii) electrochemically reduced graphene oxide, and (iii) carbon black-functionalized devices. The findings suggest that improved electron transfer kinetics and increased active surface area of the CNs, along with surface oxygen content, aid in the detection of anti-SARS-CoV-2 IgG antibodies. The novel immunosensor suggests a possible application toward monitoring of IgG antibody production in SARS-CoV-2-vaccinated patients to study immune responses, vaccine efficacy, and lifetime to meet the demands for POC analysis in resource-limited settings.

Electrochemical study of N-acetyl neuraminic acid at gold-modified screen-printed carbon electrodes for a glycoprotein sensor of SARS-CoV-2

Abstract An electrochemical sensor of SARS-CoV-2 spike glycoprotein S1 has been developed using N-acetyl neuraminic acid (Neu5Ac) as the biosensing agent based on the interaction between Neu5Ac with the N-terminal domain of spike glycoprotein S1. Gold-modified screen-printed carbon electrodes (Au-SPCEs), prepared by electrochemical deposition of gold particles on a screen-printed carbon electrode surface, were used to detect the presence of the SARS-CoV-2 from the change in the electrochemical behavior of Neu5Ac. Cyclic voltammetry of Neu5Ac in phosphate buffer solution pH 7.6 using Au-SPCEs yielded a couple of oxidation and reduction peaks at potentials of +0.6 and 0.1 V (vs Ag/AgCl), respectively. The currents of both peaks linearly increased with the Neu5Ac concentrations in the range from 0 to 200 µM. A sensitivity of 0.77 mA/mM and 0.47 mA/mM for reduction and oxidation currents were observed, respectively, with estimated detection limits (LODs) of 51.05 µM and 35.77 µM. In the presence of SARS-CoV-2 spike glycoprotein S1, these current peaks decreased. At an optimum contact time of 30 min, a linear correlation to the current change of the reduction peaks was observed for the spike glycoprotein S1 over the concentration range 0 to 0.01 µg/mL. An estimated LOD of 0.7 ng/mL could be achieved with excellent stability (2.18% RSD value for n = 10), indicating that the sensor is promising for use in real applications of SARS-CoV-2 spike glycoprotein S1 detection.

The optimization of self-assembled monolayer of thiols on screen-printed gold electrode

<span>The activated gold-modified electrode surface for self-assembled monolayer (SAM) is suitable for N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), 16-mercaptohexadecanoic acid (16-MDA), and N-hydroxysuccinimide (NHS). The various substrates could potentially be used for surface functionalization that binds to one another. Using modified screen-printed gold electrodes, 16-MDA, EDC, and NHS, we developed a self-assembling monolayer. Different 16-MDA concentrations were applied to the surface of a screen-printed electrode surface to enhance the sensitivity of the working electrode. The impact of different EDC, NHS, and 16-MDA concentrations (0.4 M, 0.8 M, 1 M, 1.2 M, 1.4 M, and 1.8 M) and incubation times between 5 and 30 minutes were examined and compared. The binding surface of the screen-printed gold electrode was characterized using the differential pulse voltammetry (DPV) technique. It has been demonstrated that the substrate concentrations at 5 minutes and 30 minutes of incubation time used to have the highest surface coverage and electron transfer rates. This concentration would be utilized in the subsequent experiment to evaluate the binding of various bacterial species. These findings suggest that the SAM in a modified screen-printed gold electrode may be functionalized to detect microorganisms.</span>

Development of an electrochemical biosensor utilizing a combined aptamer and MIP strategy for the detection of the food allergen lysozyme.

This study aims to develop a MIP-Apt-based electrochemical biosensor for the sensitive and selective determination of Lysozyme (Lyz), a food allergen. For the development of the sensor, in the first stage, modifications were made to the screen-printed electrode (SPE) surface with graphene oxide (GO) and gold nanoparticles (AuNPs) to increase conductivity and surface area. The advantages of using aptamer (Apt) and molecularly imprinted polymer (MIP) technology were combined in a single biointerface in the prepared sensing tool. Surface characterization of the biosensor was performed using scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectrometry (XPS), contact angle measurements, cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). A wide linear range from 0.001 to 100pM was obtained under optimized conditions for the determination of Lyz detection usingthe proposed MIP-Apt sensing strategy. The limit of detection (LOD) and limit of quantification (LOQ) for Lyz were 3.67 fM and 12 fM, respectively. This biosensor displays high selectivity, repeatability, reproducibility, and long storage stability towards Lyz detection. The results show that a sensitive and selective sensor fabrication is achieved compared with existing methods.

Nanomolar detection of essential amino acid in dairy products using a novel electrochemical sensor based on zinc cobaltite nanoflowers embedded porous 3D reduced graphene oxide

L-tryptophan (L-Trp) is a vital amino acid that sways neuronal function, immunity, and gut homeostasis, and its accurate detection in food samples is crucial. The aim of this study is to integrate zinc cobaltite (ZCO) nanoparticles and 3D porous reduced graphene oxide (rGO) on a screen-printed carbon electrode (SPCE) surface for building a novel electrochemical sensor to sensitively detect L-Trp in food products. A low-temperature aqueous solution method was employed in ZCO nanoflower synthesis and a hydrothermal approach was utilized to prepare 3D rGO. SEM, elemental mapping, XRD, Raman spectroscopy, XPS, and EIS characterizations were performed on the prepared nanocomposite, ZCO/3DrGO. Electrochemical experiments conducted with the cyclic and differential pulse voltammetric techniques were used for effectively assessing the catalytic power of ZCO/3DrGO/SPCE. With a low detection limit of 3 nM, high sensitivity of 19.53 μAμM−1cm−2, and a broad linear range of 0.08–5.93; 5.93–87.18 μM, the sensor demonstrated promising electrocatalytic activity towards L-Trp. Further, the reliability of the sensor was proved by analyzing its stability, repeatability, reproducibility, and selectivity towards L-Trp. The successful detection of L-Trp in dairy products (yogurt, milk, and cottage cheese) using the proposed sensor evinced its practical feasibility with high recovery of 98.16%–101.16% and low RSD of 2.8%.