Disposable Screen-printed Carbon Electrode Research Articles

Cytochrome c electrochemical detection utilizing molecularly imprinted poly(3, 4-ethylenedioxythiophene) on a disposable screen printed carbon electrode

Cytochrome c (cyt c) has been found to play a function in apoptosis in cell-free models. This work presents the creation of molecularly imprinted conducting poly(3, 4-ethylenedioxythiopene) (MIPEDOT) on the surface of a screen printed carbon electrode (SPCE) for cyt c. Cyt c was imprinted by electropolymerization due to the presence of an EDOT monomer hydrophobic functional group on SPCE, using CV to obtain highly selective materials with excellent molecular recognition ability. MIPEDOT was characterized by CV, EIS, and DPV using ferricyanide/ferrocyanide as a redox probe. Further, the characterization of the sensor was accomplished using SEM for surface morphological confirmation. Using CV, the peak current measured at the potential of +1 to −1 V (vs. Ag/AgCl) is linear in the cyt c concentration range from 1 to 1200 pM, showing a remarkably low detection limit of 0.5 pM (sensitivity:0.080 μA pM). Moreover, the applicability of the approach was successfully confirmed with the detection of cyt c in biological samples (human plasma). Similarly, our research has proven a low-cost, simple, and efficient sensing platform for cyt c detection, rendering it a viable tool for the future improvement of reliable and exact non-encroaching cell death detection.

Novel Deliberately Sensitive and Selective Tetrahydrozoline Voltammetric Sensors Integrated with a Copper Oxide Nanoparticle/Zeolite Platform.

The present study introduced a novel disposable screen-printed carbon electrodes (SPCEs) modified with copper oxide/zeolite nanostructures for eco-friendly selective differential pulse voltammetric quantification of tetrahydrozoline (THZ) in eyedrop samples and biological fluids. Modification of the electrode matrix with copper oxide nanoparticles/zeolite nanostructures (CuONPs/ZY) with their effective and synergistic electrocatalytic activity enhanced the electrode performance against electrooxidation of THZ at 0.960 V in BR at pH 9.0 with a diffusion-controlled reaction mechanism. The tentative oxidation mechanism based on molecular orbital calculations postulates the oxidation of THZ molecules through oxidation of a nitrogen atom five-membered ring and the participation of two electrons/protons in the electrode reaction. Linear calibration curves were illustrated within a wide THZ concentration range from 0.24 to 57.2 μg mL-1 recording a limit of detection (LOD) value of 0.0799 μg mL-1. The CuONPs/ZY/SPEs exhibited improved performance compared with the sole reported THZ sensor-based gold film-plated carbon paste electrodes, in addition to their high reproducibility of fabrication and measurement and prolonged shelf lifetime. Tetrahydrozoline was successfully assayed in the presence of excipients, degradation products, and chloramphenicol. The presented voltammetric sensor can be considered as an eco-friendly and reliable analytical approach for monitoring THZ residues in eye drop samples and biological fluids with high recovery compared with the official pharmacopeial analytical protocol. The presented sensors were assessed according to an EcoScale tool and also compared with the reported THZ sensor.

Rational design of microflower-like manganese-doped ultrathin cobalt hydroxide nanosheets as bifunctional electrode materials for electrochemical sensing and supercapacitor applications

Noble metal-based electrocatalyst must be replaced for electrochemical applications by earth-rich, highly active, low-cost, and highly efficient dual-function electrocatalysts. In this study, transition metal-based hydroxides (TMHs), such as 2D manganese-doped cobalt hydroxide (Mn-Co(OH)2) ultrafine nanosheets, are investigated using a very simple large-scale co-precipitation method for efficient electrochemical sensing (nitrofurantoin (NFT)) and supercapacitor (SC) applications. The characterization results show that the porous and large surface area, polycrystalline nature, confinement of the 2D orientation, and strong synergistic effect contribute to the admirable electrochemical concert of Mn-Co(OH)2 for both sensing and SC applications. CV, GCD, EIS, and DPV techniques are used to thoroughly investigate electrochemical and physicochemical properties. As an electrochemical sensor, the Mn-Co(OH)2 modified disposable screen-printed carbon electrode (Mn-Co(OH)2/SPCE) shows excellent electrochemical assets for NFT detection with broad linear ranges (0.05–219 μM and 262–641 μM), low detection limit (0.012 μM) and good anti-interference ability. The obtained specific capacitance of Mn-Co(OH)2 is 348.8 F g−1 at 1 A g−1 with a maintenance capacitance of 80.76 % after 5000 cycles at 10 A g−1. The enhanced electrochemical behavior of Mn-Co(OH)2 may be related to the fast diffusion pathways of Mn doping and the enhanced redox reaction (cobalt ions) of the flower-like ultrathin nanosheets owing to their high surface area (214 m2/g). The characterization results and electrochemical performance (sensing and SCs) provide important and noteworthy insights into the synthesis of noble metal-free and high-efficiency transparent 2D TMHs nanosheets with high performance that can be widely used in various applications.

Smartphone-based photoelectrochemical immunoassay for carcinoembryonic antigen based on BiOCl/CuBi2O4 heterojunction

Photoelectrochemical (PEC) immunoassay has been widely developed for biomarker detection, but most include heavy and expensive instruments that are not suited for portable and on-site detection. In this work, the PEC immunoassay platform for mobile phones was reported for flexible, rapid, low-cost detection of carcinoembryonic antigen (CEA). The PEC detection platform was successfully composed of disposable screen-printed carbon electrodes, a micro-electrochemical workstation, a flashlight (the excitation light source), and a smartphone with a companion software with a micro-electrochemical workstation for rapid and on-site detection of target biomarkers. In this portable smartphone-based PEC system, the S-scheme heterojunction BiOCl/CuBi2O4 was effectively excited due to the efficient electron transfer rate and excellent photocurrent response under visible light. Specifically, the sandwich-type immunoreaction for capturing target biomarkers introduced alkaline phosphatase (ALP) labeled gold nanoparticles (Au NPs). The addition of CEA increased the ascorbic acid (AA) content and enhanced the photocurrent. The proposed immunoassay presented a good linear with the logarithm of CEA concentrations range within 0.01–40 ng mL−1, and the detection limit of 3.5 pg mL−1 (S/N = 3). Therefore, the portable detection platform offered an implementable approach to the development of miniaturized and portable photoelectrochemical detectors and on-site detection technology.

Highly sensitive electrochemical volatile organic compound (Acetone) sensing based on TiO2 Nanocubes @Polyaniline nanostructure

The identification of volatile organic compounds (VOCs) as biomarkers has gained significant attention as a non-invasive point-of-care (POC) diagnostic route. VOC concentrations in the human body offer insights into pathophysiological states and aid in the detection of various diseases. This research article focuses on the synthesis of TiO2 Nanocubes (TNC) and TNC@Polyaniline nanocomposite (TNCP) via sol–gel and in-situ polymerization. Structural and morphological analyses were conducted using XRD, FESEM, and TEM techniques. Additionally, disposable screen-printed carbon electrodes (SPCE) were employed to investigate the electrochemical behaviour. The TNCP/SPCE-based VOC biomarkers exhibited exceptional performance, including a remarkably low detection limit of 0.002 µM and a superior sensitivity of 2869.76 µA mM−1cm−2 within the linear range of 1–1000 μM. These results demonstrate that TNCP holds significant potential as a superior VOC biomarker in biomedical applications, particularly for diabetic diagnosis.

Highly Efficient CeO2–CuCrO2 Composite Nanofibers Used for Electrochemical Detection of Dopamine in Biomedical Applications

Dopamine (DA) plays a crucial role in the functioning of the human central nervous system, participating in both physiological and psychological processes. It is an important research topic in biomedical science. However, we need to constantly monitor the concentration of dopamine in the body, and the sensors required for this usually require good sensitivity in order to achieve fast and accurate measurements. In this research project, a CeO2 and CuCrO2 composite nanofiber was prepared for the electrochemical detection of dopamine. Coaxial electrospinning techniques were used to prepare CeO2–CuCrO2 composite nanofibers. The characterization techniques of X-ray diffractometer (XRD), Raman, and X-ray photoelectron spectroscopy (XPS) were used to analyze the composite’s crystal structure, vibrational bonds, and elemental composition, while SEM and TEM were used to analyze the composite’s surface structure, morphology, and microstructure. The prepared nanofiber outer layer was found to have an average thickness of 70.96 nm, average fiber diameter of 192.49 nm, and an average grain size of about ~12.5 nm. The BET analysis was applied to obtain the specific surface area (25.03 m2/gm). The proposed nanofiber-decorated disposable screen-printed carbon electrode acted as a better electrochemical sensor for the detection of dopamine. Moreover, the electrocatalyst had a better limit of detection, 36 nM with a linear range of 10 to 100 μM, and its sensitivity was 6.731 μA μM−1 cm−2. In addition, the proposed electrocatalyst was successfully applied to real-time potential applications, namely, to the analysis of human urine samples in order to obtain better recovery results.

Development of interdigitated electrode based enzyme sensor for triglyceride monitoring using engineered oxygen insensitive glycerol 3-phosphate oxidase

Triglycerides (TG) are an important biomarker of various diseases including hyper-lipidemia. Current emergent requirements for TG sensing include the development of electrochemical sensors for easy, cost-effective, disposable and single use triglyceride sensors. We propose a TG sensor using an engineered glycerol 3-phosphate oxidase (GlpO) and electrochemical detection. In this study, we aimed to develop an electrochemical TG sensor based on engineering the oxidative half reaction of GlpO to be oxygen insensitive, with the employment of an interdigitated electrode (IDE). To construct an oxygen insensitive mutant GlpO, we constructed a double mutant, Met104Phe/Asp169Glu, which revealed an engineered GlpO exhibiting repressed oxidase activity, while its dye-mediated dehydrogenase activity enhanced. The electrochemical enzyme sensor was constructed for the measurement of TG based on engineered GlpO using disposable screen-printed carbon electrode (SPCE) or disposable IDE using Ru complex as the electron mediator. By using the disposable IDE sensor, a highly sensitive electrochemical sensor for Glp was achieved, with the sensitivity of 0.4 µA/mM. The improved sensitivity was 2.4 times higher than that of SPCE (0.17 µA/mM), with LOD of 0.022 mM. Furthermore, the dry sensor strip, where the double mutant GlpO and Ru complex were dropcasted and dried on the IDE, was prepared and the Glp concentration was successfully measured in the delipidized serum solution. Therefore, by the employment of the established pretreatment method for TG to release Glp, a highly sensitive, accurate, easy and rapid electrochemical measurement method for TG will be developed using constructed M104F/D169E mutant GlpO with the combination of IDE.

Point-of-care testing: a disposable label-free electrochemical CA125 and HE4 immunosensors for early detection of ovarian cancer.

Cancer antigen 125 (CA125) and human epididymal secretory protein 4 (HE4) are critical biomarkers for ovarian cancer diagnosis and progression monitoring; therefore, sensitive determination of their levels in body fluids is crucial. In recent study, label-free CA125 and HE4 immunosensors were prepared using disposable screen-printed carbon electrodes modified with reduced graphene oxide, polythionine, and gold nanoparticles for the sensitive, fast, and practical determination of CA125 and HE4. Differential pulse voltammetry, square wave voltammetry, and electrochemical impedance spectroscopy methods were used for the electrochemical determination of antigens in four different linear ranges (1-100 pg mL- 1, 0.01-10 ng mL- 1, 10-50 ng mL- 1, and 50-500 ng mL- 1). High sensitivity, low limit of detection, and limit of quantification were obtained for each linear range with a correlation coefficient above 0.99. The application stability of CA125 and HE4 immunosensors was determined as 60 days, and the storage stability was determined as 16 weeks. Immunosensors showed high selectivity in nine different antigen mixtures. The reusability of the immunosensors has been tested up to 9 cycles. The Risk of Ovarian Malignancy Algorithm score% values were calculated using the concentration of CA125 and HE4 in the blood serum and evaluated in terms of ovarian cancer risk. For the point-of-care testing, CA125 and HE4 levels at pg mL- 1 concentration were measured in blood serum samples using the developed immunosensors and a hand-held electrochemical reader in approximately 20-30s, and high recoveries were obtained. These disposable label-free immunosensors are user-friendly and can be used in point-of-care tests for rapid and practical detection of CA125 and HE4 with high selectivity, sensitivity, and repeatability.

An innovative wireless electrochemical card sensor for field-deployable diagnostics of Hepatitis B surface antigen

A wireless-based detection utilizing an innovative electrochemical card (eCard) sensor controlled by a smartphone was developed for targeting Hepatitis B surface antigen (HBsAg). A simple label-free electrochemical platform allows a convenient operation for point-of-care diagnosis. A disposable screen-printed carbon electrode was modified straightforwardly layer-by-layer with chitosan followed by glutaraldehyde, allowing a simple but effective, reproducible, and stable method for covalently immobilizing antibodies. The modification and immobilization processes were verified by electrochemical impedance spectroscopy and cyclic voltammetry. The smartphone-based eCard sensor was used to quantify HBsAg by measuring the change in current response of the [Fe(CN)6]3-/4- redox couple before and after the presence of HBsAg. Under the optimal conditions, the linear calibration curve for HBsAg was found to be 10–100,000 IU/mL with a detection limit of 9.55 IU/mL. The HBsAg eCard sensor was successfully applied to detect 500 chronic HBV-infected serum samples with satisfactory results, demonstrating the excellent applicability of this system. The sensitivity and specificity of this sensing platform were found to be 97.75% and 93%, respectively. As illustrated, the proposed eCard immunosensor offered a rapid, sensitive, selective, and easy-to-use platform for healthcare providers to rapidly determine the infection status of HBV patients.

A label-free dual immunosensor for the simultaneous electrochemical determination of CA125 and HE4 biomarkers for the early diagnosis of ovarian cancer.

The blood levels of cancer antigen 125 (CA125) and human epididymal secretory protein 4 (HE4) are measured in the diagnosis and progression monitoring of ovarian cancer (OC), and the Risk of Ovarian Malignancy Algorithm (ROMA) score% values are calculated for cancer risk assessment. For the first time, disposable dual screen-printed carbon electrodes modified with reduced graphene oxide, polythionine, and gold nanoparticles were used to fabricate label-free electrochemical dual CA125-HE4 immunosensors for the sensitive, fast, and practical simultaneous determination of CA125 and HE4. DPV and SWV methods were used to simultaneously determine antigens in two different linear ranges (1-100pgmL-1 and 1-50ngmL-1). High sensitivity, low LOD, and LOQ were obtained for two linear ranges with a correlation coefficient above 0.99. The application stability of the dual CA125-HE4 immunosensors was determined as 60days, and the storage stability was determined as 16weeks. The dual immunosensors exhibited high selectivity in eight different antigen mixtures. The reusability of the dual immunosensors has been tested up to 9 cycles. ROMA score% values for pre-menopausal and post-menopausal status were calculated using the concentration of CA125 and HE4 in the blood serum and assessing OC risk. The disposable dual immunosensors can be used in point-of-care tests for rapid and practical simultaneous determination of CA125 and HE4 with high selectivity, sensitivity, and repeatability.

Click and Detect: Versatile Ampicillin Aptasensor Enabled by Click Chemistry on a Graphene-Alkyne Derivative.

Tackling the current problem of antimicrobial resistance (AMR) requires fast, inexpensive, and effective methods for controlling and detecting antibiotics in diverse samples at the point of interest. Cost-effective, disposable, point-of-care electrochemical biosensors are a particularly attractive option. However, there is a need for conductive and versatile carbon-based materials and inks that enable effective bioconjugation under mild conditions for the development of robust, sensitive, and selective devices. This work describes a simple and fast methodology to construct an aptasensor based on a novel graphene derivative equipped with alkyne groups prepared via fluorographene chemistry. Using click chemistry, an aptamer is immobilized and used as a successful platform for the selective determination of ampicillin in real samples in the presence of interfering molecules. The electrochemical aptasensor displayed a detection limit of 1.36nM, high selectivity among other antibiotics, the storage stability of 4 weeks, and is effective in real samples. Additionally, structural and docking simulations of the aptamer shed light on the ampicillin binding mechanism. The versatility of this platform opens up wide possibilities for constructing a new class of aptasensor based on disposable screen-printed carbon electrodes usable in point-of-care devices.

Heterostructure of amorphous nickel boride anchored 2D-layered cobalt selenide nanosheets as a disposable strip sensor for detection of toxic pollutant 5-nitroquinoline in aquatic samples

The development of disposable electrode-based sensors is of great importance for the electrochemical monitoring of toxic pollutants in wastewater systems, which are often present and mutually inhibiting during sample detection. In this work, a distinctive approach was taken to use an electrochemical heterostructure sensor with chemically reduced nickel boride (Ni3B) nanoparticles anchored in two-dimensional (2D) layered cobalt selenide (CoSe) nanosheets. The prepared hybrid structure (2D CoSe/Ni3B) enables large active sites and a strong synergistic effect between nanostructures to improve the electrocatalytic activity in the detection of water pollutant 5-nitroquinoline (5-NQ) with a disposable screen-printed carbon electrode (SPCE). The 2D CoSe/Ni3B/SPCE exhibits a reduction peak potential of −0.63 V for 5-NQ, with an increased reduction peak current compared to that of other electrodes. The fabricated disposable strip electrode is suitable for the electrochemical testing of pollutants. The proposed hybrid 2D CoSe/Ni3B/SPCE disposable sensor showed a wide linear range of 0.2–378 μM and a low detection limit of 0.042 μM for 5-NQ. The developed 2D CoSe/Ni3B hybrid nanostructure-based disposable sensor efficiently detects 5-NQ in hazardous water contaminants by voltammetric analysis and showed acceptable recoveries. Based on the results, it is believed that the proposed work provides a clear idea for the fabrication of transition metal-based metal boride and selenide heterostructures with excellent catalytic activity for the electrochemical detection of various pollutants in real-time monitoring.

Ultrawide Hydrazine Concentration Monitoring Sensor Comprising Ir-Ni Nanoparticles Decorated with Multi-Walled Carbon Nanotubes in On-Site Alkaline Fuel Cell Operation.

A highly sensitive amperometric hydrazine monitoring sensor offering an ultrawide dynamic range of 5 μM to 1 M in alkaline media (e. g., 1 M KOH) was developed via co-electrodepositing iridium-nickel alloy nanoparticles (NPs) functionalized with multi-walled carbon nanotubes (Ir-Ni-MWCNTs) on a disposable screen-printed carbon electrode. The synergistic interaction of MWCNTs with Ir-Ni alloy NPs resulted in enlarged active surface area, rapid electron transfer, and alkaline media stability with an onset potential of -0.12 V (vs. Ag/AgCl) toward hydrazine oxidation. A limit of detection for hydrazine was 0.81 μM with guaranteed reproducibility, repeatability, and storage stability alongside a superb selectivity toward ethanolamine, urea, dopamine, NaBH4 , NH4 OH, NaNO2 , and Na2 CO3 . The sensor was finally applied to on-site monitoring of the carbon-free hydrazine concentration at the anode and cathode of a hydrazine fuel cell, providing more insight into the hydrazine oxidation process during cell operation.

Electrochemical Sensors Based on Au Nanoparticles Decorated Pyrene-Reduced Graphene Oxide for Hydrazine, 4-Nitrophenol and Hg2+ Detection in Water.

Monitoring hazardous chemical compounds such as hydrazine (N2H4), 4-nitrophenol (4-NP) and Hg2+ in natural water resources is a crucial issue due to their toxic effects on human health and catastrophic impact on the environment. Electrochemical nanostructured platforms integrating hybrid nanocomposites based on graphene derivatives and inorganic nanoparticles (NPs) are of great interest for such a purpose. In this work, disposable screen-printed carbon electrodes (SPCEs) have been modified with a hybrid nanocomposite formed by reduced graphene oxide (RGO), functionalized by 1-pyrene carboxylic acid (PCA), and decorated by colloidal Au NPs. These hybrid platforms have been tested for the electrocatalytic detection of N2H4 and 4-NP by differential pulse voltammetry and have been modified with an electropolymerized film of Hg2+ ions imprinted polycurcumin for the electroanalytical detection of Hg2+ by DPV. LODs, lower and in line with the lowest ones reported for state-of-the-art electrochemical sensors, integrating similar Au-graphene < nanocomposites, have been estimated. Additionally, good repeatability, reproducibility, and storage stability have been assessed, as well as a high selectivity in the presence of a 100-fold higher concentration of interfering species. The applicability of the proposed platforms for the detection of the compounds in real complex matrices, such as tap and river water samples, has been effectively demonstrated.

Electrochemical biosensor for quantitative determination of fentanyl based on immobilized cytochrome c on multi-walled carbon nanotubes modified screen-printed carbon electrodes.

Fentanyl is a powerful synthetic opioid used to treat severe pain. New administration routes toward its illegal consumption for recreational purposes pose a growing threat to public health, either due to misuse or abuse of this substance. As a result, the rapid qualitative and quantitative determination of fentanyl in biofluids is of great interest. A novel enzymatic biosensor based on adsorptive-stripping cyclic voltammetry is proposed as a cost-effective, reliable, and efficient device for fentanyl determination in urine samples. Disposable screen-printed carbon electrodes modified with multi-walled carbon nanotubes and cytochrome c were used to develop the testing platform. The electrochemical behavior of fentanyl exhibited a well-defined anodic wave around 0.66Vvs. pseudo reference electrode. The experimental conditions were optimized to obtain the best analytical response, and linear regression analysis of increasing concentration standards was applied to estimate the performance parameters. The results suggest a simple method with a wide linearity range, high sensitivity, low limits of detection (0.086μg/mL) and quantification, and satisfactory precision (2.9%RSD). The feasibility and applicability of the voltammetric approach were assessed by fentanyl-spiked urine samples by standard additions calibration curves in two levels of enrichment with an accuracy of 92% and 100%.

Simultaneous Amperometric Aptasensor Based on Diazonium Grafted Screen-Printed Carbon Electrode for Detection of CFP10 and MPT64 Biomarkers for Early Tuberculosis Diagnosis

Early diagnosis is highly crucial for life-saving and transmission management of tuberculosis (TB). Despite the low sensitivity and time-consuming issues, TB antigen detection still relies on conventional smear microscopy and culture techniques. To address this limitation, we report the development of the first amperometric dual aptasensor for the simultaneous detection of Mycobacterium tuberculosis secreted antigens CFP10 and MPT64 for better diagnosis and control of TB. The developed sensor was based on the aptamers-antibodies sandwich assay and detected by chronoamperometry through the electrocatalytic reaction between peroxidase-conjugated antibodies, H2O2, and hydroquinone. The CFP10 and MPT64 aptamers were immobilized via carbodiimide covalent chemistry over the disposable dual screen-printed carbon electrodes modified with a 4-carboxyphenyl diazonium salt. Under optimized conditions, the aptasensor achieved a detection limit of 1.68 ng mL-1 and 1.82 ng mL-1 for CFP10 and MPT64 antigens, respectively. The developed assay requires a small sample amount (5 µL) and can be easily performed within 2.5 h. Finally, the dual aptasensor was successfully applied to clinical sputum samples with the obtained diagnostic sensitivity (n = 24) and specificity (n = 13) of 100%, respectively, suggesting the readiness of the developed assay to be used for TB clinical application.

Low-cost, disposable sensors based on polystyrene waste and V–TiO2 microneedle composites for diagnosis of renal malfunction and muscle dystrophy

A compact, cost-efficient, and disposable electrochemical creatinine sensor is designed using disposable screen-printed carbon electrodes coated with a composite receptor based on polystyrene foam (i.e., packaging material waste) and anatase-phase V–TiO2 microneedles. The electrochemical sensor is capable of monitoring 1–25 μM creatinine in a standard redox solution at 7.4 pH and room temperature. The electrochemical measurements are performed with a portable potentiostat using an android smartphone. Cyclic voltammetry and differential pulse voltammetry characteristics are measured to monitor the concentration of creatinine. The sensor exhibits excellent creatinine sensitivity of 30.95 ± 1.73 μA/cm2⸳μM and an extremely low threshold detection limit of 184 nM creatinine.

A sandwich-like configuration with a signal amplification strategy using a methylene blue/aptamer complex on a heterojunction 2D MoSe2/2D WSe2 electrode: Toward a portable and sensitive electrochemical alpha-fetoprotein immunoassay.

Liver cancer is one of the most common global health problems that features a high mortality rate. Alpha-fetoprotein (AFP) is a potential liver cancer biomarker for the diagnosis of liver cancer. The quantitative detection of AFP at an ultratrace level has important medical significance. Using the reaction of the antibody-antigen pair in an immunosensor enables the sensitive and selective AFP assay. Finding a strategy in signal generation and amplification is challenging to fabricate new sensitive electrochemical immunosensors for AFP detection. This study demonstrates the construction of a simple, reliable, and label-free immunosensor for the detection of AFP on a smart phone. Exfoliated two-dimensional (2D) molybdenum diselenide (MoSe2) and 2D tungsten diselenide (WSe2) were employed to modify the disposable screen-printed carbon electrode (SPCE) to use as the electrochemical platform, which is affixed to a small potentiostat connected to a smart phone. The modified electrode offers antibody immobilization and allows detection of AFP via an immunocomplex forming a sandwich-like configuration with the AFP-corresponding aptamer. A heterojunction 2D MoSe2/2D WSe2 composite improves the SPCE's reactivity and provides a large surface area and good adsorption capacity for the immobilizing antibodies. The signal generation for the immunosensor is from the electrochemical response of methylene blue (MB) intercalating into the aptamer bound on the electrode. The response for the proposed sandwich-like immunosensor is proportional to the AFP concentration (1.0-50,000 pg ml-1). The biosensor has potential for the development of a simple and robust point-of-care diagnostic platform for the clinical diagnosis of liver cancer, achieving a low limit of detection (0.85 pg ml-1), high sensitivity, high selectivity, good stability, and excellent reproducibility.

Synthetic Biology Toolbox, Including a Single-Plasmid CRISPR-Cas9 System to Biologically Engineer the Electrogenic, Metal-Resistant Bacterium Cupriavidus metallidurans CH34.

Cupriavidus metallidurans CH34 exhibits extraordinary metabolic versatility, including chemolithoautotrophic growth; degradation of BTEX (benzene, toluene, ethylbenzene, xylene); high resistance to numerous metals; biomineralization of gold, platinum, silver, and uranium; and accumulation of polyhydroxybutyrate (PHB). These qualities make it a valuable host for biotechnological applications such as bioremediation, bioprocessing, and the generation of bioelectricity in microbial fuel cells (MFCs). However, the lack of genetic tools for strain development and studying its fundamental physiology represents a bottleneck to boosting its commercial applications. In this study, inducible and constitutive promoter libraries were built and characterized, providing the first comprehensive list of biological parts that can be used to regulate protein expression and optimize the CRISPR-Cas9 genome editing tools for this host. A single-plasmid CRISPR-Cas9 system that can be delivered by both conjugation and electroporation was developed, and its efficiency was demonstrated by successfully targeting the pyrE locus. The CRISPR-Cas9 system was next used to target candidate genes encoding type IV pili, hypothesized by us to be involved in extracellular electron transfer (EET) in this organism. Single and double deletion strains (ΔpilA, ΔpilE, and ΔpilAE) were successfully generated. Additionally, the CRISPR-Cas9 tool was validated for constructing genomic insertions (ΔpilAE::gfp and ΔpilAE::λPrgfp). Finally, as type IV pili are believed to play an important role in extracellular electron transfer to solid surfaces, C. metallidurans CH34 ΔpilAE was further studied by means of cyclic voltammetry using disposable screen-printed carbon electrodes. Under these conditions, we demonstrated that C. metallidurans CH34 could generate extracellular currents; however, no difference in the intensity of the current peaks was found in the ΔpilAE double deletion strain when compared to the wild type. This finding suggests that the deleted type IV pili candidate genes are not involved in extracellular electron transfer under these conditions. Nevertheless, these experiments revealed the presence of different redox centers likely to be involved in both mediated electron transfer (MET) and direct electron transfer (DET), the first interpretation of extracellular electron transfer mechanisms in C. metallidurans CH34.

Capacitive nanobiosensing of β-1,4-galactosyltransferase-V colorectal cancer biomarker

β-1,4-GalT-V is an enzyme with glycosyltransferase activity that glycosylates proteins and synthesizes the lactosylceramide sphingolipid in the Golgi apparatus. Colorectal cancer (CRC) tumor cells produce these biomolecules in high concentrations concerning normal cells, releasing them into the blood serum. Hence, β-1,4-GalT-V glycoprotein has emerged as a promising CRC biomarker, and its detection opens opportunities for the diagnosis/prognosis of CRC. We report the first capacitive nanobiosensor for the detection of β-1,4-GalT-V based on disposable screen-printed carbon electrodes (SPCEs) nanostructured with gold nanorods (AuNRs) and the Prussian blue (PrB) redox-active compound. AuNRs increased the SPCE surface area for antibody immobilization by physical adsorption and promoted the deposition of PrB onto the SPCE surface. Antibody-glycoprotein molecular biorecognition event onto the SPCE/AuNRs/PrB surface perturbed the PrB redox density-of-states and hence redox capacitance (Cμ̅). We found that changes in the inverse of Cμ̅ correlated well with the increasing concentrations of β-1,4-GalT-V in the linear range from 50 to 400 fM with a sensitivity of 1.5 µF fM-1 cm-2 and a limit of detection of 20 fM. The nanobiosensor detected the β-1,4-GalT-V glycoprotein from raw human serum samples with high specificity, ultrasensitivity, and reagentless label-free electrochemical transduction. The nanobiosensor could be applied as a CRC diagnosis/prognosis tool in a decentralized setting, with minimal patient sample manipulation and rapid response.