Smartphone-Controlled Aptasensor for Voltammetric Detection of Patulin in Apple Juice.

Patulin (PAT) is a mycotoxin that adversely affects the health of humans and animals. PAT can be particularly found in products such as apples and apple juice and can cause many health problems if consumed. Therefore, accurate and sensitive determination of PAT is very important for food quality and human and animal health. A voltammetric aptasensor was introduced in this study for PAT determination while measuring the changes at redox probe signal. The limit of detection (LOD) was found to be 0.18 pg/mL in the range of 1-104 pg/mL of PAT in buffer medium under optimum experimental conditions. The selectivity of the PAT aptasensor against ochratoxin A, fumonisin B1 and deoxynivalenol mycotoxins was examined and it was found that the aptasensor was very selective to PAT. PAT determination was performed in an apple juice medium for the first time by using a smartphone-integrated portable device, and accordingly, an LOD of 0.47 pg/mL was achieved in diluted apple juice medium. A recovery range of 91.24-93.47% was obtained for PAT detection.

Carbon Nanofiber-Ionic Liquid Nanocomposite Modified Aptasensors Developed for Electrochemical Investigation of Interaction of Aptamer/Aptamer-Antisense Pair with Activated Protein C.

Selective and sensitive detection of human activated protein C (APC) was performed herein by using carbon nanofiber (CNF) and ionic liquid (IL) composite modified pencil graphite electrode (PGE) and electrochemical impedance spectroscopy (EIS) technique. A carbon nanomaterial-based electrochemical aptasensor was designed and implemented for the first time in this study for the solution-phase interaction of DNA-Apt with its cognate protein APC as well as APC inhibitor aptamer-antidote pair. The applicability of this assay developed for the determination of APC in fetal bovine serum (FBS) and its selectivity against different proteins (protein C, thrombin, bovine serum albumin) was also examined. CNF-IL modified aptasensor specific to APC provided the detection limit as 0.23 µg/mL (equal to 3.83 nM) in buffer medium and 0.11 µg/mL (equal to 1.83 nM) in FBS. The duration of the proposed assay from the point of electrode modification to the detection of APC was completed within only 55 min.

Implementation of 3D printing technologies to electrochemical and optical biosensors developed for biomedical and pharmaceutical analysis.

Three-dimensional (3D) printing technology has been applied in many areas. In recent years, new generation biosensors have been emerged with the progress on 3D printing technology (3DPT). Especially in the development of optical and electrochemical biosensors, 3DPT provides many advantages such as low cost, easy to manufacturing, being disposable and allow point of care testing. In this review, recent trends in the development of 3DPT based electrochemical and optical biosensors with their applications in the field of biomedical and pharmaceutical are examined. In addition, the advantages, disadvantages and future opportunities of 3DPT are discussed.

Recent Progress on Optical Biosensors Developed for Nucleic Acid Detection Related to Infectious Viral Diseases.

Optical biosensors have many advantages over traditional analytical methods. They enable the identification of several biological and chemical compounds directly, instantly, and without the need of labels. Their benefits include excellent specificity, sensitivity, compact size, and low cost. In this review, the main focus is placed on the nucleic acid-based optical biosensor technologies, including colorimetric, fluorescence, surface plasmon resonance (SPR), Evanescent-Wave Optical, Fiber optic and bioluminescent optical fibre. The fundamentals of each type of biosensor are briefly explained, and particular emphasis has been placed on the achievements which have been gained in the last decade on the field of diagnosis of infectious viral diseases. Concluding remarks concerning the perspectives of further developments are discussed.

Zip Nucleic Acid-Based Genomagnetic Assay for Electrochemical Detection of microRNA-34a.

Zip nucleic acid (ZNA)-based genomagnetic assay was developed herein for the electrochemical detection of microRNA-34a (miR-34a), which is related to neurological disorders and cancer. The hybridization between the ZNA probe and miR-34a target was performed in the solution phase; then, the resultant hybrids were immobilized onto the surface of magnetic beads (MBs). After magnetic separation, the hybrids were separated from the surface of MBs and then immobilized on the surface of pencil graphite electrodes (PGEs). In the case of a full-match hybridization, the guanine oxidation signal was measured via the differential pulse voltammetry (DPV) technique. All the experimental parameters that influenced the hybridization efficiency (i.e., hybridization strategy, probe concentration, hybridization temperature, etc.) were optimized. The cross-selectivity of the genomagnetic assay was tested against two different miRNAs, miR-155 and miR-181b, individually as well as in mixture samples. To show the applicability of the ZNA-based genomagnetic assay for miR-34a detection in real samples, a batch of experiments was carried out in this study by using the total RNA samples isolated from the human hepatocellular carcinoma cell line (HUH-7).

Aptasensor for Impedimetric Detection of Lysozyme.

The impedimetric detection of a protein, lysozyme (LYS), was carried out herein by aptamer-immobilized single-use pencil graphite electrodes (PGEs). The aptamer was immobilized onto electrochemically activated surface of electrode without using any chemical agents, or any types of nanomaterials. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques were applied to analyze the electrochemical behavior of unmodified PGE and aptamer immobilized PGE. The interaction of aptamer with its target protein, LYS, was then investigated by EIS. The limit of detection for LYS was found to be 1.44 µg/mL (equals to 100.65 nM). The developed aptasensor specific to LYS presented high selectivity against to bovine serum albumin and thrombin.

Diagnostic kit based on halloysite nanoclay-ionic liquid nanocomposite modified electrode for electrochemical determination of cancer biomarker.

Nucleic acid hybridization is occurred between the selective single-stranded nucleic acid sequence and its target sequence, which is one of the essential procedure for electrochemical detection of nucleic acid. microRNA-21 (miRNA-21) is known as a biomarker in various cancers. The determination of miRNA-21 was attained through by hybridization of inosine substituted miRNA-21 specific DNA probe (Pinosine) with its target miRNA-21. In this study, the surface of pencil graphite electrode (PGE) was firstly modified with halloysite nanoclay-ionic liquid (HNT/IL) nanocomposite. The characterization of surface was performed by Scanning Electron Microscope (SEM) images and Energy Dispersive X-Ray Analysis (EDX) analysis, and the differences at surface modifications were also shown by electrochemical methods with electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). For sensitive and selective determination of miRNA-21, Pinosine and target miRNA concentration, immobilization and hybridization time were optimized by using HNT/IL modified PGE in combination with differential pulse voltammetry (DPV). The detection limit was achieved as 0.17 µg/mL (equals to 23.69 nM) in the linear range of 0.25-2 µg/mL miRNA-21. The selectivity of voltammetric method based on HNT/IL-PGE developed for miRNA-21 was examined in the presence of mismatch (MM) and non-complementary (NC) sequences. Because miRNA-21 is over-expressed in cancer cells, it has been tested in total RNA samples isolated from cancer cell line (breast cancer cell line, MCF-7). In the total RNA samples obtained from MCF-7, the detection limit was calculated as 0.28 µg/mL in the linear range of 1-4 µg/mL. Besides, the healthy cell line (human embryonic kidney cell line, HEK-293) was used as a control group and the results obtained by MCF-7 total RNA samples were compared to the results using HEK-293 total RNA samples in terms of miRNA-21 level.

Electrochemical DNA biosensors developed for the monitoring of biointeractions with drugs: a review.

The interaction of drugs with DNA is important for the discovery of novel drug molecules and for understanding the therapeutic effects of drugs as well as the monitoring of side effects. For this reason, many studies have been carried out to investigate the interactions of drugs with nucleic acids. In recent years, a large number of studies have been performed to electrochemically detect drug-DNA interactions. The fast, sensitive, and accurate results of electrochemical techniques have resulted in a leading role for their implementation in this field. By means of electrochemical techniques, it is possible not only to demonstrate drug-DNA interactions but also to quantitatively analyze drugs. In this context, electrochemical biosensors for drug-DNA interactions have been examined under different headings including anticancer, antiviral, antibiotic, and central nervous system drugs as well as DNA-targeted drugs. An overview of the studies related to electrochemical DNA biosensors developed for the detection of drug-DNA interactions that were reported in the last two decades in the literature is presented herein along with their applications and they are discussed together with their future perspectives.

Low-Cost High-Resolution Potentiostat for Electrochemical Detection of Nucleic Acids and Biomolecular Interactions.

A handheld USB-powered instrument developed for the electrochemical detection of nucleic acids and biomolecular interactions is presented. The proposed instrument is capable of scanning ± 2.25 V while measuring currents up to ±10 mA, with a minimum current resolution of 6.87 pA. Therefore, it is suitable for nucleic acid sensors, which have high background currents. A low-cost microcontroller with an on-chip 16-bit analog-to-digital converter, 12-bit digital-to-analog converter, and a built-in USB controller were used to miniaturize the system. The offset voltages and gain errors of the analog peripherals were calibrated to obtain a superior performance. Thus, a similar performance to those of the market-leader potentiostats was achieved, but at a fraction of their cost and size. The performance of the application of this proposed architecture was tested successfully and was found to be similar to a leading commercial device through a clinical application in the aspects of the detection of nucleic acids, such as calf thymus ssDNA and dsDNA, and their interactions with a protein (BSA) by using single-use graphite electrodes in combination with the differential pulse voltammetry technique.

Impedimetric Detection Based on Label-Free Immunoassay Developed for Targeting Spike S1 Protein of SARS-CoV-2.

After the COVID-19 pandemic started all over the world, great importance was placed on the development of sensitive and selective bioanalytical assays for the rapid detection of the highly pathogenic SARS-CoV-2 virus causing COVID-19 disease. In this present work, an impedimetric immunosensor was developed and applied for rapid, reliable, sensitive and selective detection of the SARS-CoV-2 S1 protein. To detect the SARS-CoV-2 virus, targeting of the spike S1 protein was achieved herein by using S1 protein-specific capture antibody (Cab-S1) immobilized screen-printed electrode (SPE) in combination with the electrochemical impedance spectroscopy (EIS) technique. With the impedimetric immunosensor, the detection limit for S1 protein in buffer medium was found to be 0.23 ng/mL (equal to 23.92 amol in 8 µL sample) in the linear concentration range of S1 protein from 0.5 to 10 ng/mL. In the artificial saliva medium, it was found to be 0.09 ng/mL (equals to 9.36 amol in 8 µL sample) in the linear concentration range of S1 protein between 0.1 and 1 ng/mL. The selectivity of the impedimetric immunosensor toward S1 protein was tested against influenza hemagglutinin antigen (HA) in the buffer medium as well as in artificial saliva.

Amperometric immunosensor developed for sensitive detection of SARS-CoV-2 spike S1 protein in combined with portable device.

In this present study, an amperometric immunosensor was developed based on disposable screen-printed carbon electrode (SPCE) for specific and sensitive detection of SARS-CoV-2 S1 protein. Anti-SARS-CoV-2 S1 monoclonal antibody was firstly immobilized onto the electrode surface. Then, the sandwich complex was formed by addition of S1 protein, secondary antibody and HRP-IgG, respectively. Chronoamperometry measurements were done in the presence of TMB mediator and the detection of SARS-CoV-2 S1 protein was performed by using 10 µL sample. The limit of detection (LOD) was found to be 0.19 ng/mL (equals to 24.7 amol in 10 µL sample) in the linear range of 0.5-10 ng/mL obtained in buffer medium. The applicability of this assay was investigated in the linear range of 0.5-3 ng/mL S1 protein in artificial saliva medium with the LOD as 0.13 ng/mL (equals to 16.9 amol in 10 µL sample). The selectivity study was examined in the presence of Hemagglutinin antigen (HA) in both mediums; buffer and artificial saliva while resulting with the successful discrimination between S1 protein and HA. The one of ultimate goals of our study is to present the possible implementation of this assay to point of care (POC) analysis. Under this aim, this assay was performed in combination with a portable device that is the commercial electrochemical analyzer. Amperometric detection of S1 protein in the range of 0.5-5 ng/mL was also successfully performed in artificial saliva medium with a resulting LOD as 0.15 ng/mL (equals to 19.5 amol in 10 µL sample). In addition, a selectivity study was similarly carried out by portable device.

Graphene-Oxide and Ionic Liquid Modified Electrodes for Electrochemical Sensing of Breast Cancer 1 Gene.

Graphene-oxide and ionic liquid composite-modified pencil graphite electrodes (GO-IL-PGEs) were developed and used as a sensing platform for breast cancer 1 (BRCA1) gene detection. The characterization of GO-IL modified electrodes was executed by scanning electron microscopy (SEM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The nucleic-acid hybridization was monitored by a differential pulse voltammetry (DPV) technique by directly measuring the guanine oxidation signal without using any indicator. The effects of the IL concentration, the probe concentration, and the hybridization time were optimized to the biosensor response. The limit of detection (LOD) was calculated in the concentration range of 2-10 µg/mL for the BRCA1 gene and found to be 1.48 µg/mL. The sensitivity of the sensor was calculated as 1.49 µA mL/µg cm2. The developed biosensor can effectively discriminate the complementary target sequence in comparison to a three-base-mismatched sequence or the non-complementary one.

Impedimetric detection of miRNA biomarkers using paper-based electrodes modified with bulk crystals or nanosheets of molybdenum disulfide.

Paper-based electrodes modified with molybdenum disulfide (MoS2) in the form of bulk crystals or exfoliated nanosheets were developed and used as a biosensing platform for the impedimetric detection of miRNAs (miRNA-155 and miRNA-21) related to early diagnosis of lung cancer. For this purpose, MoS2 crystals or nanosheets were used for the modification of the working electrode area of paper-based platform for the first time in this study. The proposed assay offers sensitive and selective detection of microRNAs by electrochemical impedance spectroscopy (EIS) technique. The entire assay, both the electrode modification and the miRNA detection being completed in 30 min and a single sample droplet (5 µL) was enough to cover working electrode area which enabled analysis in low sample volumes. The limits of detection (LOD) for miRNA-21 and miRNA-155 were calculated both in buffer and fetal bovine serum media. It is found that the LOD is varying between 1 and 200 ng/mL. In comparison to nanosheets, a larger electroactive surface area was obtained with bulk MoS2 resulting in lower LOD values on miRNA detection. The paper-based electrodes showed high specificity towards their target sequences. Moreover, they effectively discriminated single base mismatched non-target sequences. The advantages of these MoS2 paper based electrodes include high sensitivity, and low-cost provide great potential for improved monitoring of miRNA biomarkers even in artificial serum media.

Preparation of Surface Plasmon Resonance Aptasensor for Human Activated Protein C Sensing.

Nucleic acid aptamers are an emerging class of artificial ligands and have recently gained attention in several areas. Here we report the design of a surface plasmon resonance (SPR) aptasensor for highly sensitive and selective sensing of human activated protein C (APC). First, DNA aptamer (DNA-Apt) specific for APC is complexed with N-methacryloyl-L-cysteine (MAC) monomer. Then, 2-hydroxyethyl methacrylate (HEMA) and cyanamide are mixed with the DNA-Apt/MAC complex. The SPR aptasensor is characterized by atomic force microscopy, ellipsometry, and contact angle measurements. Selectivity of SPR aptasensor is carried out in the presence of myoglobin (Myb), hemoglobin (Hb), and bovine serum albumin (BSA). Limit of detection (LOD) and limit of quantification (LOQ) values are 1.5 ng mL-1 and 5.2 ng mL-1, respectively. DNA-Apt SPR aptasensor performance for APC detection is also examined in artificial plasma.

Detection of Senecionine in Dietary Sources by Single-Use Electrochemical Sensor.

Pyrrolizidine alkaloids (PAs) are produced by plants as secondary compounds that are the most widely distributed natural toxins. There have been many cases of human toxicity caused by consumption of toxic plant species, as herbal teas and grain or grain products contaminated with PA-containing seeds have been reported. Companies that produce dried spices and tea leaves should examine the PA level in their products. For the first time in the literature, a simple and inexpensive electrochemical assay based on a single-use sensor was introduced for quantitative determination of senecionine (SEN) in the most frequently contaminated food sources. SEN was immobilized on a pencil graphite electrode surface by the passive adsorption technique. Differential pulse voltammetry (DPV) was used to evaluate the oxidation signal of SEN, which was observed to be around +0.95 V. The oxidation signal was specific to the SEN in the sample, and the current value was proportional to its concentration. The selectivity of our assay was also tested in the presence of other similar PAs such as intermedine, lycopsamine, and heliotrine. The detection limit is calculated by developed assay and found to be 5.45 µg/mL, which is an acceptable concentration value of SEN occurring at toxic levels for consumers. As an application of the developed sensor in food products, the electrochemical detection of SEN was successfully performed in flour and herbal tea products.

Paper-Based Electrochemical Biosensors for Voltammetric Detection of miRNA Biomarkers Using Reduced Graphene Oxide or MoS2 Nanosheets Decorated with Gold Nanoparticle Electrodes.

Paper-based biosensors are considered simple and cost-efficient sensing platforms for analytical tests and diagnostics. Here, a paper-based electrochemical biosensor was developed for the rapid and sensitive detection of microRNAs (miRNA-155 and miRNA-21) related to early diagnosis of lung cancer. Hydrophobic barriers to creating electrode areas were manufactured by wax printing, whereas a three-electrode system was fabricated by a simple stencil approach. A carbon-based working electrode was modified using either reduced graphene oxide or molybdenum disulfide nanosheets modified with gold nanoparticle (AuNPs/RGO, AuNPs/MoS2) hybrid structures. The resulting paper-based biosensors offered sensitive detection of miRNA-155 and miRNA-21 by differential pulse voltammetry (DPV) in only 5.0 µL sample. The duration in our assay from the point of electrode modification to the final detection of miRNA was completed within only 35 min. The detection limits for miRNA-21 and miRNA-155 were found to be 12.0 and 25.7 nM for AuNPs/RGO and 51.6 and 59.6 nM for AuNPs/MoS2 sensors in the case of perfectly matched probe-target hybrids. These biosensors were found to be selective enough to distinguish the target miRNA in the presence of single-base mismatch miRNA or noncomplementary miRNA sequences.

Electrochemical Investigation of Curcumin-DNA Interaction by Using Hydroxyapatite Nanoparticles-Ionic Liquids Based Composite Electrodes.

Hydroxyapatite nanoparticles (HaP) and ionic liquid (IL) modified pencil graphite electrodes (PGEs) are newly developed in this assay. Electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and cyclic voltammetry (CV) were applied to examine the microscopic and electrochemical characterization of HaP and IL-modified biosensors. The interaction of curcumin with nucleic acids and polymerase chain reaction (PCR) samples was investigated by measuring the changes at the oxidation signals of both curcumin and guanine by differential pulse voltammetry (DPV) technique. The optimization of curcumin concentration, DNA concentration, and the interaction time was performed. The interaction of curcumin with PCR samples was also investigated by gel electrophoresis.

Cobalt Phthalocyanine-Ionic Liquid Composite Modified Electrodes for the Voltammetric Detection of DNA Hybridization Related to Hepatitis B Virus.

In this study, cobalt phthalocyanine (CoPc) and ionic liquid (IL) modified pencil graphite electrodes (PGEs) were designed and implemented to detect sequence-selective DNA hybridization related to the Hepatitis B virus (HBV). The surface characterization of CoPc-IL-PGEs was investigated by scanning electron microscopy (SEM), and the electrochemical behavior of electrodes were studied by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques. The voltammetric detection of hybridization was investigated by evaluating the guanine oxidation signal, measured by differential pulse voltammetry (DPV) technique. The implementation of our biosensor to serum samples was also examined using fetal bovine serum (FBS). The detection limit was established as 0.19 µg/mL in phosphate buffer solution (PBS) (pH 7.40) and 2.48 µg/mL in FBS medium. The selectivity of our assay regarding HBV DNA hybridization in FBS medium was tested in the presence of other DNA sequences. With this aim, the hybridization of DNA probe with non-complementary (NC) or mismatched DNA sequence (MM), or in the presence of mixture samples containing DNA target NC (1:1) or DNA target MM (1:1), was studied based on the changes in guanine signal.

Paper-based electrode assemble for impedimetric detection of miRNA.

In the present work, a paper-based electrode assemble was developed and implemented to detect target microRNA 155 (miRNA 155) via electrochemical impedance spectroscopy (EIS) measurements. In this concept, gold nanoparticles (AuNPs) modified paper based electrode assemble system (AuNP-PE) was designed, and characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and EIS measurements. The impedimetric detection of miRNA 155 was performed by measuring the fractional change at the charge transfer resistance (Rct). The detection limits were found as 33.8 nM in PBS and 93.4 nM in fetal bovine serum (FBS) medium, respectively. The selectivity of the proposed assay was tested against to non-complementary (NC) and mismatch (MM) miRNA sequences in the presence of mixture sample containing miRNA:NC (1:1) and miRNA:MM (1:1) in PBS (pH 7.40) or FBS. The analytical performance and the selectivity of impedimetric biosensor were also tested in FBS.

Hybrid nanoflowers modified pencil graphite electrodes developed for electrochemical monitoring of interaction between Mitomycin C and DNA.

In the present study, biocompatible hybrid nanoflowers (NFs) were synthesized by amino acids (glycine, l-lysine) via a simple, rapid and cost-effective methods. NFs were characterized by using FT-IR, Raman spectroscopy, XPS, SEM and EDX techniques. Modified pencil graphite electrode (PGE) surfaces with well-defined NFs were developed to electrochemical monitoring of calf thymus double stranded DNA (ctdsDNA) using differential pulse voltammetry (DPV) for the first time. SEM, EDX, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) methods were used to characterize the surfaces obtained after modification. In comparison to l-lysine NFs (LNFs-PGE), glycine NFs (GNFs-PGE) exhibited a higher sensitivity performance towards the oxidation of guanine moiety signals. The interaction time between anticancer drug Mitomycin C (MC) and ctdsDNA was aslo investigated with GNFs-PGE.