Recent Prospects of Carbonaceous Nanomaterials-Based Laccase Biosensor for Electrochemical Detection of Phenolic Compounds.

Phenolic compounds (PhCs) are ubiquitously distributed phytochemicals found in many plants, body fluids, food items, medicines, pesticides, dyes, etc. Many PhCs are priority pollutants that are highly toxic, teratogenic, and carcinogenic. Some of these are present in body fluids and affect metabolism, while others possess numerous bioactive properties such as retaining antioxidant and antimicrobial activity in plants and food products. Therefore, there is an urgency for developing an effective, rapid, sensitive, and reliable tool for the analysis of these PhCs to address their environmental and health concern. In this context, carbonaceous nanomaterials have emerged as a promising material for the fabrication of electrochemical biosensors as they provide remarkable characteristics such as lightweight, high surface: volume, excellent conductivity, extraordinary tensile strength, and biocompatibility. This review outlines the current status of the applications of carbonaceous nanomaterials (CNTs, graphene, etc.) based enzymatic electrochemical biosensors for the detection of PhCs. Efforts have also been made to discuss the mechanism of action of the laccase enzyme for the detection of PhCs. The limitations, advanced emerging carbon-based material, current state of artificial intelligence in PhCs detection, and future scopes have also been summarized.

Graphitic Carbon Nitride-Wrapped Metal-free PoPD-Based Biosensor for Xanthine Detection.

A metal-free, enzymatic biosensor was developed using graphitic carbon nitride (g-C3N4)-wrapped poly-ortho-phenylenediamine (PoPD) for the determination of xanthine (Xn). Field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction confirmed the successful formation of the PoPD, g-C3N4 nanosheets and PoPD@g-C3N4 nanocomposite. Furthermore, the electrochemical behavior of the biosensor was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The prepared enzyme electrode exhibited maximum response at pH 7.5 with a response time of 5 s, and its sensitivity was 5.798 µAM-1. The nanocomposite shows exceptional sensing capabilities for detecting Xn, having a wide linear range from 1 nM to 1 µM with a relatively low detection limit of 0.001 nM. The biosensor shows good stability (4 weeks) and reproducibility and can detect the presence of Xn from other interfering analytes. Validation of the biosensor with real samples obtained from Rohu (Labeo rohita) fish shows that the fabricated biosensor has the requisite potential to be used for Xn detection in meat samples.

Development of conducting cellulose paper for electrochemical sensing of procalcitonin.

An electrochemical paper-based sensor was developed for the detection of bacterial infection (BI)-specific biomarker procalcitonin (PCT). Reduced graphene oxide-gold nanoparticles (rGO-AuNP) and poly(3,4-ethylene dioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) were synthesized and were fabricated to a disposable, portable, and inexpensive cellulose fiber paper (CFP) substrate. rGO-AuNP-PEDOT:PSS nanocomposite-modified conductive paper-based biosensing platform was efficaciously fabricated by a constant and simple coating procedure. rGO-AuNP-PEDOT:PSS nanocomposite-modified conductive paper electrode was found to provide a sensitive and conductive substrate for PCT detection. The presence of rGO-AuNP-PEDOT:PSS nanocomposite on CFP substate was investigated by Fourier transform infrared spectrometry, field emission scanning electron microscopy, ultraviolet-visible spectroscopy, and X-ray diffraction studies. The electrochemical behavior of rGO-AuNP-PEDOT:PSS @CFP surface was studied with impedance spectroscopy, cyclic voltammetry, and chronoamperometry techniques. This low-cost paper-based biosensor has a linear range for PCT of 1 × 103 to 6 × 107 fg mL-1. This developed sensor exhibited good reproducibility with a relative standard deviation (RSD) of about 3.7%. The proposed CFP-based biosensor has been proven as an accelerated simple point-of-care (POC) exploratory approach for early PCT diagnosis in inadequate areas with limited production facilities, computational techniques, and highly skilled experts.

Highly Sensitive Enzymatic Biosensor Based on Polyaniline-Wrapped Titanium Dioxide Nanohybrid for Fish Freshness Detection.

Herein, we envisage the fabrication of a highly sensitive enzymatic electrochemical biosensor for selective detection of xanthine (Xn) using xanthine oxidase (XOs) immobilized polyaniline-wrapped titanium dioxide (PANI@TiO2) nanohybrid as a sensing platform. The PANI@TiO2 nanohybrid was synthesized via chemical polymerization using ammonium per sulfate as an oxidant. Various microscopic, spectroscopic, and electrochemical techniques have been utilized to confirm the electrophoretic deposition of the PANI and PANI@TiO2 nanohybrids on to indium tin oxide (ITO) coated glass substrate. The fabricated XOs/PANI@TiO2/ITO electrode exhibits enhanced electron transfer kinetics with an electron transfer rate constant of 0.904 cm s-1. The electrochemical results show that the fabricated biosensor can detect Xn in the concentration range 1-100 µM, with a limit of detection of 0.1 µM (S/N = 3) and a response time of 10 s. The validation of the biosensors has been conducted using real samples obtained from the rohu (Labeo rohita) fish. The proposed biosensor can be a reliable analytical tool for determining Xn concentration in commercial fish and meat samples.

Highly sensitive electrochemical detection of cancer biomarker based on anti-EpCAM conjugated molybdenum disulfide grafted reduced graphene oxide nanohybrid.

An ultrasensitive, electrochemical biosensor has been fabricated by utilizing molybdenum disulfide (MoS2) grafted reduced graphene oxide (MoS2@rGO) nanohybrid as a sensing platform. Biomolecular-assisted synthetic method was adopted to synthesize MoS2@rGO nanohybrid, where L-cys was used to reduce GO. The MoS2@rGO nanohybrid exhibits improved electrochemical performance when it has been electrophoretically deposited onto the indium tin oxide (ITO) coated glass substrate. Further, epithelialcell adhesion moleculeantibodies (anti-EpCAM) specific to cancer biomarker has been covalently immobilized on the MoS2@rGO/ITO electrodes for label-free detection of EpCAM. Electrochemical results confirm that anti-EpCAM/MoS2@rGO/ITO based biosensor can detect EpCAM in the concentration range of 0.001-20 ng mL-1 with a detection limit of 44.22 fg mL-1 (S/N = 3). The biosensor's excellent analytical performance has been attributed to the efficient immobilization of EpCAM antibodies on the MoS2@rGO surface, which results in high specificity for EpCAM antigen. The fabricated biosensor showed good selectivity, reproducibility, and stability. The successful detection of EpCAM antigen in spiked samples (human saliva, serum and urine) makes this platform an alternative method for early screening of cancer biomarker.

Gold nanomaterials for optical biosensing and bioimaging.

Gold nanoparticles (AuNPs) are highly compelling nanomaterials for biomedical studies due to their unique optical properties. By leveraging the versatile optical properties of different gold nanostructures, the performance of biosensing and biomedical imaging can be dramatically improved in terms of their sensitivity, specificity, speed, contrast, resolution and penetration depth. Here we review recent advances of optical biosensing and bioimaging techniques based on three major optical properties of AuNPs: surface plasmon resonance, surface enhanced Raman scattering and luminescence. We summarize the fabrication methods and optical properties of different types of AuNPs, highlight the emerging applications of these AuNPs for novel optical biosensors and biomedical imaging innovations, and discuss the future trends of AuNP-based optical biosensors and bioimaging as well as the challenges of implementing these techniques in preclinical and clinical investigations.

Electrochemical biosensor for the epithelial cancer biomarker EpCAM based on reduced graphene oxide modified with nanostructured titanium dioxide.

An electrochemical immunosensor has been fabricated for the early determination of epithelial cell adhesion molecules (EpCAM, tumor biomarker) antigen using reduced graphene oxide (rGO) modified with nanostructured titanium dioxide (TiO2). The hydrothermally synthesized rGO@TiO2 nanocomposite has been electrophoretically deposited on indium tin oxide (ITO) coated glass substrate, and the deposition was confirmed using various spectroscopic, microscopic, and electrochemical techniques. The fabricated rGO@TiO2/ITO electrode shows improved electron transfer kinetics with an electron transfer rate constant of 1.93 × 10-7 cm·s-1. Furthermore, the rGO@TiO2/ITO electrodes were used for the covalent immobilization of monoclonal EpCAM antibodies. Electrochemical determination of the EpCAM cancer biomarker is achieved using differential pulse voltammetry by scanning the potential from - 0.4 to 0.8 V with an amplitude of 50 mV. The rGO@TiO2-based biosensor shows high sensitivity (3.24 µA·mL·ng-1·cm-2), wide detection range (0.01 ng·mL-1 to 60 ng·mL-1), and low detection limit (0.0065 ng·mL-1, S/N = 3). The fabricated biosensor is highly stable and regenerable and has been successfully applied to the determination of EpCAM in spiked human serum samples. Graphical abstract.

Nanomaterial-Modified Conducting Paper: Fabrication, Properties, and Emerging Biomedical Applications.

The emerging demand for wearable, lightweight portable devices has led to the development of new materials for flexible electronics using non-rigid substrates. In this context, nanomaterial-modified conducting paper (CP) represents a new concept that utilizes paper as a functional part in various devices. Paper has drawn significant interest among the research community because it is ubiquitous, cheap, and environmentally friendly. This review provides information on the basic characteristics of paper and its functionalization with nanomaterials, methodology for device fabrication, and their various applications. It also highlights some of the exciting applications of CP in point-of-care diagnostics for biomedical applications. Furthermore, recent challenges and opportunities in paper-based devices are summarized.

An impedimetric biosensor based on electrophoretically assembled ZnO nanorods and carboxylated graphene nanoflakes on an indium tin oxide electrode for detection of the DNA of Escherichia coli O157:H7.

Aminopropyltrimethoxysilane (APTMS)-functionalized zinc oxide (ZnO) nanorods and carboxylated graphene nanoflakes (c-GNF) were used in a composite that was electrophoretically deposited on an indium tin oxide (ITO) coated glass substrate. The modified ITO electrodes were characterized using various microscopic and spectroscopic techniques which confirm the deposition of the APTMS-ZnO/c-GNF composite. The electrodes have been used for the covalent immobilization of an Escherichia coli O157:H7 (E. coli)-specific DNA prob. Impedimetric studies revealed that the gene sensor displays linear response in a wide range of target DNA concentration (10-16 M to 10-6 M) with a detection limit of 0.1 fM. The studies on the cross-reactivity to other water-borne pathogens show that the bioelectrode is highly specific. Graphical abstractSchematic illustration for fabrication of nucleic acid biosensor for E. coli DNA detection using an ITO electrode modified with siloxane-functionalized zinc oxide (ZnO) nanorods and carboxylated graphene nanoflakes (c-GNFs).

Highly efficient Polyaniline-MoS2 hybrid nanostructures based biosensor for cancer biomarker detection.

In this work, polyaniline nanospindles have been synthesized using iron oxide as sacrificial template. These nanospindles were utilized for the fabrication of PANI-MoS2 nanoflower architectures via hydrothermal route. The electrostatic interaction between PANI and MoS2 improves the conductivity and provides more direct paths for charge transportation. SEM, TEM, XRD, Raman Spectroscopy techniques were employed to explore the crystal structure, and morphological properties of the PANI-MoS2 nanocomposite. Furthermore, an electrochemical biosensing platform based on PANI-MoS2 nanocomposite was fabricated for the specific detection of chronic myelogenous leukemia (CML) by using electrochemical impedance spectroscopy technique. The binding interactions between the pDNA/PANI-MoS2/ITO bioelectrode and target DNA sequence were also studied. The biosensor exhibits high sensitivity and wide detection range (10-6  M to 10-17  M) of target DNA with low detection limit (3 × 10-18  M). Additionally, the specificity studies of the genosensor with various target DNA sequences (complementary, noncomplementary and one base mismatch) and real samples analysis of CML shows its potential for clinical diagnostics.

Electrochemical genosensor based on template assisted synthesized polyaniline nanotubes for chronic myelogenous leukemia detection.

This work reports a facile approach to synthesize polyaniline nanotubes (PANI-NT) by using manganese oxide as sacrificial templates. This template assisted polyaniline nanotubes (t-PANI-NT) were utilized as electrode material after deposition onto the indium tin oxide (ITO) coated glass substrates by using the electrophoretic technique. The structural, morphological and electrochemical characterizations of the t-PANI-NT show relatively better results compared to chemically synthesized PANI-NT (c-PANI-NT). Moreover, the t-PANI-NT/ITO electrode exhibits improved electron transfer coefficient (α = 0.63) and charge transfer rate constant (ks = 0.05912 s-1) in comparison to c-PANI-NT/ITO electrode (α = 0.56 and ks = 0.06548 s-1). The obtained t-PANI-NT/ITO electrodes have been further immobilized with biotinylated DNA sequence, specific to chronic myelogenous leukemia (CML) by using avidin-biotin as a cross-linking agent. Electrochemical impedance spectroscopy studies revealed that the genosensor displays linearity in wide range of target DNA concentration (10-6 to 10-16 M) with an outstanding differentiation ability and low detection limit of 10-16 M. The experimental results of this highly sensitive and specific genosensor with clinical samples of CML positive patients and control negative patients indicate its potential for clinical diagnostics.

Controlled deposition of functionalized silica coated zinc oxide nano-assemblies at the air/water interface for blood cancer detection.

We report results of the studies relating to controlled deposition of the amino-functionalized silica-coated zinc oxide (Am-Si@ZnO) nano-assemblies onto an indium tin oxide (ITO) coated glass substrate using Langmuir-Blodgett (LB) technique. The monolayers have been deposited by transferring the spread solution of Am-Si@ZnO stearic acid prepared in chloroform at the air-water interface, at optimized pressure (16 mN/m), concentration (10 mg/ml) and temperature (23 °C). The high-resolution transmission electron microscopic studies of the Am-Si@ZnO nanocomposite reveal that the nanoparticles have a microscopic structure comprising of hexagonal assemblies of ZnO with typical dimensions of 30 nm. The surface morphology of the LB multilayer observed by scanning electron microscopy shows uniform surface of the Am-Si@ZnO film in the nanometer range (<80 nm). These electrodes have been utilized for chronic myelogenous leukemia (CML) detection by covalently immobilizing the amino-terminated oligonucleotide probe sequence via glutaraldehyde as a crosslinker. The response studies of these fabricated electrodes carried out using electrochemical impedance spectroscopy show that this Am-Si@ZnO LB film based nucleic acid sensor exhibits a linear response to complementary DNA (10(-6)-10(-16) M) with a detection limit of 1 × 10(-16) M. This fabricated platform is validated with clinical samples of CML positive patients and the results demonstrate its immense potential for clinical diagnosis.

Gold nanoparticle decorated graphene sheet-polypyrrole based nanocomposite: its synthesis, characterization and genosensing application.

We report herein the synthesis of gold nanoparticle (GNP) decorated-graphene sheets (GO-GNPs) using the template of graphene oxide (GO) by a one-pot solution-based method. A polypyrrole-GO decorated GNP nanocomposite (GO-GNP/PPY) has been electropolymerized using a potentiodynamic method on an indium tin oxide (ITO) coated glass substrate. The as-synthesized nanocomposites are characterized by transmission electron microscopy, energy dispersive X-ray spectroscopy, scanning electron microscopy, thermogravimetric analysis, Fourier transform infrared and Raman spectroscopy. It has been found that GNPs of ca. 13 nm are uniformly dispersed on the surface of GO, and have a high electrochemically active surface area. The surface morphology studies show that PPY structure changes from nanoflowers to nanostars and then to nanosheets with an increase in the scan rate (20-200 mV s(-1)). The prepared GO-GNP/PPY/ITO electrode was further used as a genosensor, where the electrochemical response was measured using methylene blue (MB) as a redox indicator. The genosensor shows a response time of 60 s with high sensitivity (1 × 10(-15) M) and linearity (1 × 10(-15)-1 × 10(-6) M) with the correlation coefficient of 0.9975.

Nanostructuring of hierarchical 3D cystine flowers for high-performance electrochemical immunosensor.

Here, we report a simple and reproducible method for large scale fabrication of novel flower and palm-leaf like 3D cystine microstructures (CMs) with high uniformity having a size of 50 µm and 10 µm respectively, through a facile aqueous solution route as a function of pH and concentration. In a proof-of-concept study, the 3D CMs have been further explored to fabricate a label-free high-performance electrochemical immunosensor by immobilizing monoclonal antibodies. Electrochemical methods were employed to study the stepwise modification of the system and the electronic transduction for the detection. The fabricated immunosensor design demonstrates high performance with enhanced sensitivity (4.70 cfu ml(-1)) and linear sensing range from 10 to 3 x 10(9) cfu ml(-1) a long shelf-life (35 days) and high selectivity over other bacterial pathogens. The enhanced performance originates from a novel nanostructuring in which the CMs provide higher surface coverage for the immobilization of antibodies providing excellent electronic/ionic conductivity which result in the enhanced sensitivity.

Cationic poly(lactic-co-glycolic acid) iron oxide microspheres for nucleic acid detection.

Herein, we envisage the possibility of preparing stable cationic poly(lactic-co-glycolic acid) (PLGA) microspheres encapsulating the iron oxide nanoparticles (IONPs; 8-12 nm). The IONPs are incorporated into PLGA in organic phase followed by microsphere formation and chitosan coating in aqueous medium via nano-emulsion technique. The average size of the microspheres, as determined by dynamic light scattering are about 310 nm, while the zeta potential for the composite remains near 35 mV at pH 4.0. These microspheres are electrophoretically deposited onto indium tin oxide (ITO)-coated glass substrate used as cathode and parallel platinum plate as the counter electrode. This platform is utilized to fabricate a DNA biosensor, by immobilizing a probe sequence specific to Escherichia coli. The bioelectrode shows a surface-controlled electrode reaction with the electron transfer coefficient (α) of 0.64 and charge transfer rate constant (k(s)) of 61.73 s(-1). Under the optimal conditions, this biosensor shows a detection limit of 8.7 × 10(-14) M and is found to retain about 81% of the initial activity after 9 cycles of use.

Chitosan encapsulated quantum dots platform for leukemia detection.

We report results of the studies relating to electrophoretic deposition of nanostructured composite of chitosan (CS)-cadmium-telluride quantum dots (CdTe-QDs) onto indium-tin-oxide coated glass substrate. The high resolution transmission electron microscopic studies of the nanocomposite reveal molecular level coating of the CdTe-QDs with CS molecules in the colloidal dispersion medium. This novel composite platform has been explored to fabricate an electrochemical DNA biosensor for detection of chronic myelogenous leukemia (CML) by immobilizing amine terminated oligonucleotide probe sequence containing 22 base pairs, identified from BCR-ABL fusion gene. The results of differential pulse voltammetry reveal that this nucleic acid sensor can detect as low as 2.56 pM concentration of complementary target DNA with a response time of 60s. Further, the response characteristics show that this fabricated bioelectrode has a shelf life of about 6 weeks and can be used for about 5-6 times. The results of experiments conducted using clinical patient samples reveal that this sensor can be used to distinguish CML positive and the negative control samples.

Microstructured cystine dendrites-based impedimetric sensor for nucleic acid detection.

We report results of the studies relating to the fabrication and characterization of novel biosensing electrode by covalent immobilization of DNA onto microstructural cystine (Cys) prepared by acoustic cavitation method. The TEM investigations of these structures reveal transformation of microstructured Cys from nanorods to dendritic structure under optimum conditions. The Cys dendrites (denCys) have been investigated by XRD, FT-IR, and SEM studies. These biosensing electrodes have been fabricated by immobilization of Escherichia coli (E. coli)-specific DNA probe onto the dendritic cystine. The results of the electrochemical impedance spectroscopy studies reveal that this nucleic acid sensor exhibits linear response to cDNA in the concentration range of 10(-6) to 10(-14) M with response time of 30 min. The biosensing characteristics show that the fabricated E. coli sensor can be reused about 4 times and is stable for ∼4 weeks. The studies on cross-reactivity of the sensor for other water-borne pathogens like Salmonella typhimurium, Neisseria meningitides, and Klebsiella pneumonia reveal specificity of the bioelectrode for E. coli detection.