Determination of the total antioxidant capacity of the Chinese tea based on a novel "peroxidase/zirconium phosphonate"composite electrochemical sensor.

In this work, a novel zirconium phosphonate (ZrPR1R2) was prepared by decorating both the aminoethoxy- group (R1) and the carboxypropyl- group (R2) on the zirconium phosphate layers in order to manipulate further the immobilization of the peroxidase (POD), and an antioxidant biosensor with higher sensitivity was constructed by dropping the POD/ZrPR1R2 composite onto the glassy carbon electrode surface. The activity of the POD/ZrPR1R2 composite was detected by Uv-vis spectra. The direct electrochemical behavior, the electrocatalytic response to dissolved oxygen and hydrogen peroxide, as well as the ability to detect total antioxidant capacity in tea sample were investigated by the methods of cyclic voltammetry. The results indicated that the immobilization of POD in ZrPR1R2 nanosheets matrix enhanced the enzymatic activity, and achieved the fast and direct electron transfer between POD and glassy carbon electrode. Moreover, the POD/ZrPR1R2 composite modified electrode show the electrocatalytic response to hydrogen peroxide in the linear range of 8.8×10-8 to 8.8×10-7 mol L-1, with the detection limit of 3.3×10-8 mol L-1. Attributing to the sensitive response to dissolved oxygen, the total antioxidant capacity can be detected directly in the real tea water by this POD/ZrPR1R2 composite modified electrode.

A highly selective and sensitive sensor for promethazine based on molecularly imprinted interface coated Au/Sn bimetal nanoclusters functionalized acupuncture needle microelectrode.

Promethazine (PMZ) is an effective antihistamine that is used as a nerve tranquilizer to treat mental disorders. However, drug abuse causes harm to the human body and also pollutes the environment to a certain extent. Therefore, it is crucial to develop a highly selective and sensitive biosensor for PMZ determination. An acupuncture needle (AN) was used as an electrode in 2015, and further research on the electrode's essence in electrochemistry is needed. In this work, a sensor based on a surface imprinted film coordinated Au/Sn biometal was first fabricated on AN via electrochemistry. The obtained cavities showed complementary and suitable sites for "N atom" electron transfer through the phenyl ring structure in promethazine, which is rigorous for the configuration near the interface. Under the optimal conditions, MIP/Au/Sn/ANE exhibits a good linear relationship in the range of 0.5 µM-500 µM, and the detection limit (LOD) is 0.14 µM (S/N = 3). The sensor exhibits good repeatability, stability, and selectivity and can be successfully used to analyze and detect PMZ in human serum and environmental water. The findings are scientifically significant for AN electrochemistry and the sensors have potential for in vivo medicamentosus monitoring in the future.

A graphitic nano-onion/molybdenum disulfide nanosheet composite as a platform for HPV-associated cancer-detecting DNA biosensors.

An electrochemical DNA sensor that can detect human papillomavirus (HPV)-16 and HPV-18 for the early diagnosis of cervical cancer was developed by using a graphitic nano-onion/molybdenum disulfide (MoS2) nanosheet composite. The electrode surface for probing DNA chemisorption was prepared via chemical conjugation between acyl bonds on the surfaces of functionalized nanoonions and the amine groups on functionalized MoS2 nanosheets. The cyclic voltammetry profile of an 1:1 nanoonion/MoS2 nanosheet composite electrode had an improved rectangular shape compared to that of an MoS2 nanosheet elecrode, thereby indicating the amorphous nature of the nano-onions with sp2 distancing curved carbon layers that provide enhanced electronic conductivity, compared to MoS2 nanosheet only. The nanoonion/MoS2 sensor for the DNA detection of HPV-16 and HPV-18, respectively, was measured at high sensitivity through differential pulse voltammetry (DPV) in the presence of methylene blue (MB) as a redox indicator. The DPV current peak was lowered after probe DNA chemisorption and target DNA hybridization because the hybridized DNA induced less effective MB electrostatic intercalation due to it being double-stranded, resulting in a lower oxidation peak. The nanoonion/MoS2 nanosheet composite electrodes attained higher current peaks than the MoS2 nanosheet electrode, thereby indicating a greater change in the differential peak probably because the nanoonions enhanced conductive electron transfer. Notably, both of the target DNAs produced from HPV-18 and HPV-16 Siha and Hela cancer cell lines were effectively detected with high specificity. The conductivity of MoS2 improved by complexation with nano-onions provides a suitable platform for electrochemical biosensors for the early diagnosis of many ailments in humans.

Electrochemical Detection of ompA Gene of C. sakazakii Based on Glucose-Oxidase-Mimicking Nanotags of Gold-Nanoparticles-Doped Copper Metal-organic Frameworks.

The present work developed an electrochemical genosensor for the detection of virulence outer membrane protein A (ompA, tDNA) gene of Cronobacter sakazakii (C. sakazakii) by exploiting the excellent glucose-oxidase-mimicking activity of copper Metal-organic frameworks (Cu-MOF) doped with gold nanoparticle (AuNPs). The signal nanotags of signal probes (sDNA) that biofunctionalized AuNPs@Cu-MOF (sDNA-AuNPs@Cu-MOF) were designed using an Au-S bond. The biosensor was prepared by immobilization capture probes (cDNA) onto an electrodeposited AuNPs-modified glassy carbon electrode (GCE). AuNPs@Cu-MOF was introduced onto the surface of the GCE via a hybridization reaction between cDNA and tDNA, as well as tDNA and sDNA. Due to the enhanced oxidase-mimicking activity of AuNPs@Cu-MOF to glucose, the biosensor gave a linear range of 1.0 × 10-15 to 1.0 × 10-9 mol L-1 to tDNA with a detection limit (LOD) of 0.42 fmol L-1 under optimized conditions using differential pulse voltammetry measurement (DPV). It can be applied in the direct detection of ompA gene segments in total DNA extracts from C. sakazakii with a broad linear range of 5.4-5.4 × 105 CFU mL-1 and a LOD of 0.35 CFU mL-1. The biosensor showed good selectivity, fabricating reproducibility and storage stability, and can be used for the detection of ompA gene segments in real samples with recovery between 87.5% and 107.3%.

Label-free electrochemical immunosensor for Ochratoxin a detection in coffee samples.

Ochratoxin A (OTA) is a nephrotoxic and carcinogenic mycotoxin frequently found in coffee, which directly impacts human health and the economy of many countries. For this reason, there has been a growing need for simple and sensitive tools for the on-site detection of this mycotoxin. In this study, we developed a label-free impedimetric immunosensor to detect OTA. The biosensor was built on a thin-film gold electrode evaporated on glass substrtes, modified with a self-assembled cysteamine monolayer and anti-OTA antibodies. Atomic force microscopy and Microspectroscopy RAMAN confirmed the successful functionalization of the electrodes. The biosensor performance was evaluated by electrochemical impedance spectroscopy and the measurements indicated a linear relationship between the change in the impedance values and the OTA concentration in the range from 0.5 to 100 ng mL-1 with a limit of detection of 0.15 ng mL-1. The biosensor was highly selective and did not suffer matrix interference when analyzed in coffee samples. Furthermore, considering the small sample volumes, the short time required for analysis, and the possibility of miniaturization, the developed biosensor represents a promising analytical device for on-site coffee quality analyses.

A "peptide-target-aptamer" electrochemical biosensor for norovirus detection using a black phosphorous nanosheet@Ti3C2-Mxene nanohybrid and magnetic covalent organic framework.

Norovirus (NoV) is a major foodborne pathogen responsible for acute gastroenteritis epidemics, and establishing a robust detection method for the timely identification and monitoring of NoV contamination is of great significance. In this study, a peptide-target-aptamer sandwich electrochemical biosensor for NoV was fabricated using Au@BP@Ti3C2-MXene and magnetic Au@ZnFe2O4@COF nanocomposites. The response currents of the electrochemical biosensor were proportional to the NoV concentrations ranging from 0.01-105 copies/mL with a detection limit (LOD) of 0.003 copies/mL (S/N = 3). To our best knowledge, this LOD was the lowest among published assays to date, due to the specific recognition of the affinity peptide and aptamer for NoV and the outstanding catalytic activity of nanomaterials. Furthermore, the biosensor showed excellent selectivity, anti-interference performance, and satisfactory stability. The NoV concentrations in simulative food matrixes were successfully detected using the constructed biosensor. Meanwhile, NoV in stool samples was also successfully quantified without complex pretreatment. The designed biosensor had the potential to detect NoV (even at a low level) in foods, clinical samples, and environmental samples, providing a new method for NoV detection in food safety and diagnosing foodborne pathogens.

Evaluating the antioxidant activity of secondary metabolites of endophytic fungi from Hypericum perforatum L. by an electrochemical biosensor based on AuNPs/AC@CS composite.

Due to the variety and activity of secondary metabolites of endophytic fungi (SMEF) from medicinal plants, and the operation cumbersome of existing methods for evaluating the activity, there is urgent to establish a simple, efficient and sensitive evaluation and screening technology. In this study, the prepared chitosan functionalized activated carbon (AC@CS) composite as the electrode substrate material was used to modify glassy carbon electrode (GCE), and the gold nanoparticles (AuNPs) was deposited on AC@CS/GCE by cyclic voltammetry (CV). A ds-DNA/AuNPs/AC@CS/GCE electrochemical biosensor for evaluating the antioxidant activity of SMEF from Hypericum perforatum L. (HP L.) was fabricated using the method of layer by layer assembly. The experimental conditions affecting the evaluation results of the biosensor were optimized by square wave voltammetry (SWV) using Ru(NH3)63+ as the probe, and the antioxidant activity of various SMEF from HP L. was evaluated by the proposed biosensor. Meanwhile, the results of the biosensor were also verified by UV-vis. According to the optimized experimental results, the biosensors had a high levels of oxidative DNA damage at pH 6.0 and Fenton solution system with Fe2+ to OH- ratio of 1:3 for 30 min. Among the crude extracts of SMEF from roots, stems and leaves of HP L., the crude extracts from stems presents a high antioxidant activity, but it was weaker than l-ascorbic acid. This result was consistent with the evaluation results of UV-vis spectrophotometric method, also the fabricated biosensor presents high stability and sensitivity. This study not only provides a novel, convenient and efficient way for rapid evaluating the antioxidant activity of a wide variety of SMEF from HP L., but also provides a novel evaluation strategy for the SMEF from medicinal plants.

Molecularly imprinted miniature electrochemical biosensor for SARS-CoV-2 spike protein based on Au nanoparticles and reduced graphene oxide modified acupuncture needle.

Accurate detection of SARS-CoV-2 spike (SARS-CoV-2-S) protein is of clinical significance for early diagnosis and timely treatment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Herein, a surface molecularly imprinted miniature biosensor was fabricated. Au nanoparticles (AuNPs), reduced graphene oxide (rGO), poly(methylene blue)/poly(ionic liquids) and poly(ionic liquids) were successively electrodeposited onto the pinpoint of an acupuncture needle (AN). The molecularly imprinted miniature biosensor was obtained after the template of SARS-CoV-2-S protein was removed, which could be used for sensitive detection of SARS-CoV-2-S protein. The linear range and limit of detection (LOD) were 0.1 âˆ¼ 1000 ng mL-1 and 38 pg mL-1, respectively, which were superior to other molecularly imprinted biosensors previously reported. The developed miniature biosensor also exhibited high specificity and stability. The reliability of the biosensor was evaluated by the detection of SARS-CoV-2-S protein in clinical serum samples.

AChE-based electrochemical biosensor for pesticide detection in vegetable oils: matrix effects and synergistic inhibition of the immobilized enzyme.

Enzyme-based electrochemical biosensors have been widely deployed for the detection of a range of contaminants in different food products due to their significant advantages over other (bio)sensing techniques. Nevertheless, their performance is greatly affected by the sample matrix itself or by the matrix they are presented with in pretreated samples, both of which can impact the accuracy as well as the sensitivity of the measurements. Therefore, and in order to acquire reliable and accurate measurements, matrix effects and their influence on sensor performance should be taken into consideration. Herein, acetylcholinesterase (AChE)-modified electrochemical sensors were employed for the detection of pesticides in vegetable oils. Sensor interrogation with pretreated oil samples, spiked with carbofuran, revealed the inhibitory potential of the extracted matrix varies between different types of vegetable oil and their fatty acid content. In addition, synergies between the extracted matrix from different types of vegetable oils and the carbamate pesticide, carbofuran, were observed, which led to significant deviations of the sensor's performance from its anticipated behavior in buffered solution. Taking the aforementioned into consideration, appropriate calibration curves for each type of vegetable oil were drafted, which allowed for the highly reproducible determination of different pesticide concentrations in pretreated real samples. Collectively, a better understanding of AChE inhibition by single or multiple contaminants present in vegetable oils was gained, which can find many applications in numerous fields, ranging from sensor development to the design of new pesticides and medicinal products.

Black phosphorus nanosheets/poly(allylamine hydrochloride) based electrochemical immunosensor for the selective detection of human epididymis protein 4.

In this work, an electrochemical immunosensor based on black phosphorus nanosheets (BPNS)/poly(allylamine hydrochloride) (PAH) nanocomposite modified glassy carbon electrode was developed for the detection of ovarian cancer biomarker HE4. PAH has been applied to retain BPNS in its original honeycomb structure and to anchor biomolecules electrostatically on the transducer surface. The as synthesized nanocomposite was characterized by zeta potential analysis, scanning electron microscopy, x-ray photoelectron spectroscopy, transmission electron microscopy, high-resolution transmission electron microscopy. Subsequently, the performance of the electrochemical immunosensor was evaluated through cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. Under the optimal condition, the developed electrochemical immunosensor permitted to detect HE4 with a linear range of 0.1-300 ng ml-1and a detection limit of 0.01 ng ml-1. The developed sensor exhibited good selectivity and specificity to HE4 with negligible interference effect from common biomolecules like bovine serum albumin, lysozyme, protamine, glucose, fructose, hemoglobin and fetal bovine serum. Further, practical application of developed electrochemical immunosensor was demonstrated in spiked human serum which showed satisfactory recovery percentages.

Smartphone-based magneto-immunosensor on carbon black modified screen-printed electrodes for point-of-need detection of aflatoxin B1 in cereals.

Considering the complexities and speed of modern food chains, there is an increasing demand for point-of-need detection of food contaminants, particularly highly regulated chemicals and carcinogens such as aflatoxin B1. We report a user-friendly smartphone-based magneto-immunosensor on carbon black modified electrodes for point-of-need detection of aflatoxin B1 in cereals. For buffered analyte solutions and a corn extract sample, the assay demonstrated a low limit of detection of 13 and 24 pg/mL, respectively. The assay was also highly reproducible, exhibiting mean relative standard deviations of 3.7% and 4.0% for the buffered analyte and corn extract samples. The applicability of the assay was validated on the basis of EU guidelines and the detection capability was lower than or equal to 2 µg/kg, which is the EU maximum residue limit for aflatoxin B1 in cereals. False-positive and false-negative rates were less than 5%. Additionally, an open-source android application, AflaESense, was designed to provide a simple interface that displays the result in a traffic-light-type format, thus minimizing user training and time for data analysis. AflaESense was used for smartphone-based screening of spiked corn samples containing aflatoxin B1 (0.1, 2, and 10 ng/mL), and naturally contaminated corn containing 0.15 ng aflatoxin B1/mL. The measured values were in close agreement with spiked concentrations (r2 = 0.99), with recovery values ranging between 80 and 120%. Finally, contaminated samples correctly triggered a red alert while the non-contaminated samples led to the display of a green color of AflaESense. To the best of our knowledge, this is the first smartphone-based electrochemical system effective for screening samples for contamination with aflatoxin B1.

A universal biosensor utilizing bacteria-initiated in situ growth of electroactive polymers for bacteria-related hazards detection.

Here, an ultrasensitive and highly selective electrochemical biosensor is engineered by integrating bacteria-initiated click chemistry with in situ growth of electroactive polymers. Leveraging the unique copper-binding redox pathway of bacteria to reduce CuII to CuI, CuI-catalyzed click chemistry is initiated and high-density electroactive ferrocenyl polymers are subsequently generated and efficiently grafted on biosensing interface by potentiostatic electrochemical atom transfer radical polymerization that greatly enhances the sensitivity of electro-analytical performance. A good linearity between electrochemical signal and the logarithm of Staphylococcus aureus and Escherichia coli concentration over the range from 102 to 107 CFU/mL is obtained with detection limits down to 4 and 6 CFU/mL, respectively. In order to further expand the applicability and universality of the sensor, bacterial magnetic separation section is supplemented into the system. With the help of aptamer-based magnetic preseparation section, selective detection of target bacteria with great anti-interference is achieved in complex real samples. Moreover, this biosensor can be applied in convenient antibiotics residue detection and rapid drug resistance analysis by merely substitution of recognition element or preincubation of bacteria with different anti-bacteria drugs. Thus, after further expansion of bacterial magnetic separation section or simple replacement of the originally identification element, a universal biosensor including bacteria analysis system and antibiotics detection system with excellent analytical performance is constructed. It provides new insight into the aspects of bacteria-related hazards detection that could not only reduce the detriment caused by bacterial contamination, but also guide antibiotic rational usage and help to control the emergence of multidrug-resistant bacteria.

Tailored Biosensors for Drug Screening, Efficacy Assessment, and Toxicity Evaluation.

Biosensors have been flourishing in the field of drug discovery with pronounced developments in the past few years. They facilitate the screening and discovery of innovative drugs. However, there is still a lack of critical reviews that compare the merits and shortcomings of these biosensors from a pharmaceutical point of view. This contribution presents a critical and up-to-date overview on the recent progress of tailored biosensors, including surface plasmon resonance, fluorescent, photoelectrochemical, and electrochemical systems with emphasis on their mechanisms and applications in drug screening, efficacy assessment, and toxicity evaluation. Multiple functional nanomaterials have also been incorporated into the biosensors. Representative examples of each type of biosensors are discussed in terms of design strategy, response mechanism, and potential applications. In the end, we also compare the results and summarize the major insights gained from the works, demonstrating the challenges and prospects of biosensors-assisted drug discovery.

Assay for TET1 activity and its inhibitors screening with signal amplification by both nanoparticles and Ru(III) redox recycling.

Ten-eleven translocation protein 1 (TET1) is one member of TET proteins family which plays a key role in dynamic DNA methylation-demethylation process. Herein, a novel biosensor was constructed for TET1 detection and its inhibitors screening utilizing restriction digestion of endonuclease enzyme MspI. Half-methylated oligonucleotide (5mC DNA) was used as target and Ru(NH3)63+ as electrochemical signal probe. After the treatment by TET1 and T4 ß-glucosyltransferase (T4 ß-GT), target oligonucleotide would not be recognized and digested. If there was no TET1, the target would be digested and the response of biosensor decreased greatly. The current difference of biosensor with and without the incubation with TET1 was therefore dependent on the concentration of TET1. To increase sensitivity of the biosensor, nanostructured film at electrode surface and nanoparticles modified oligonucleotides were employed as signal amplification elements for Ru(NH3)63+ recycling. Finally, this biosensor showed high performance with a wide linear range of TET1 concentration from 3.5-21 ng/µL and a low detection limit of 0.33 ng/µL, which is superior to other existing methods. The inhibition effects of Bobcat339 on TET1 was successfully proved by our biosensor with an IC50 of 38 µM. Not only that, but the feasibility of the biosensor for inhibitors screening was evaluated and further confirmed by other compounds including two anticancer drugs and three active ingredients of traditional Chinese medicine.

Electrochemical Trans-Channel Assay for Efficient Evaluation of Tumor Cell Invasiveness.

Efficiently assessing the invasive capability of tumor cells is critical both for the research and treatment of cancer. Here, we report a novel method called the electrochemical trans-channel assay for efficient evaluation of tumor cell invasiveness. A bioinspired extracellular matrix degradation model (EDM) has been first fabricated on a porous anodic alumina (PAA) membrane to construct the electrochemical apparatus. Upon contacting the invasive tumor cells, invasive capability can be sensitively evaluated by the degree of EDM impairment, which is recorded by the electrochemical trans-channel ionic currents in a label-free manner. Compared to the most commonly used trans-well migration method, this assay can be accomplished in an efficient way that is significantly faster (20 min) and more convenient. Besides, quantitation can also be realized for monitoring the invasion process, which cannot be achieved by other currently used methods. Our proposed electrochemical trans-channel assay method has shown a synergistic effect for the evaluation of tumor cell invasiveness, providing a promising method for clinical assessment or prognostic applications of tumor metastasis.

Enzymatic Electroanalytical Biosensor Based on Maramiellus colocasiae Fungus for Detection of Phytomarkers in Infusions and Green Tea Kombucha.

In this work, we developed an enzymatic voltammetric biosensor for the determination of catechin and gallic acid in green tea and kombucha samples. The differential pulse voltammetry (DPV) methodology was optimized regarding the amount of crude enzyme extract, incubation time in the presence of the substrates, optimal pH, reuse of the biosensor, and storage time. Samples of green tea and kombucha were purchased in local markets in the city of Goiânia-GO, Brazil. High performance liquid chromatography (HPLC) and Folin-Ciocalteu spectrophotometric techniques were performed for the comparison of the analytical methods employed. In addition, two calibration curves were made, one for catechin with a linear range from 1 to 60 µM (I = -0.152 * (catechin) - 1.846), with a detection limit of 0.12 µM and a quantification limit of 0.38 µM and one for gallic acid with a linear range from 3 to 60 µM (I = -0.0415 * (gallic acid) - 0.0572), with a detection limit of 0.14 µM and a quantification limit of 0.42 µM. The proposed biosensor was efficient in the determination of phenolic compounds in green tea.

Electrochemical determination of hantavirus using gold nanoparticle-modified graphene as an electrode material and Cu-based metal-organic framework assisted signal generation.

An electrochemical biosensor was prepared for nucleic acid-based hantavirus detection using a Cu-based metal-organic framework (CuMOF) as a signal tag. The CuMOF was synthesized by the solvothermal method and then covalently bonded with signal DNA (sDNA) probes. The Au nanoparticles and reduced graphene oxide composite were deposited on the electrode surface by electroreduction as support substrate and was then functionalized with capture DNA (cDNA) probes by self-assembly. Through the complementary base pairing, the target DNA (tDNA) fragment of hantavirus hybridized with the cDNA and the sDNA in a sandwich-type format. The tDNA was detected according to the current signal of the CuMOF catalyzed reaction using o-phenylenediamine as redox substrate. The peak current of the biosensor at - 0.55 V increased linearly in proportion to the logarithmic value of the tDNA concentration from 10-15 to 10-9 mol/L, with a detection limit of 0.74 × 10-15 mol/L. Moreover, the proposed biosensor was successfully applied to detect hantavirus and was able to distinguish hantavirus from other arboviruses.

Versatile electrochemical biosensor based on bi-enzyme cascade biocatalysis spatially regulated by DNA architecture.

The regulation of biocatalytic cascades in microenvironments for high performance and extended applications is still challenging. Herein, we develop a rolling circle amplification (RCA)-based one-pot method to prepare the micron-sized DNA flowers (DFs), which achieve the co-encapsulation and spatial regulation of bi-enzyme molecules, glucose oxidase (GOx) and horseradish peroxidase (HRP). In this system, GOx and HRP are integrated into the DFs simultaneously during RCA with the bridging of magnesium between enzyme residues and phosphate backbones on DFs. The cascade of GOx/HRP is regulated with the formation of highly ordered and hydrogen-bonded water environment in the cavity of DFs, resulting in an enhanced cascade catalytic efficiency compared with that in homogeneous solution. Moreover, the high density of DNA scaffold ensures the encapsulation of GOx/HRP with high efficiency. Accordingly, a glucose electrochemical biosensor with amplified signal response is fabricated using the as-prepared GOx/HRP DFs as biosensing interface, realizing sensitive detection of glucose. Further, through designing the complementary sequence of aptamer into the programmable circular template of RCA, the bi-enzyme co-encapsulated DFs are versatilely applied to sensitive and selective detection of cancerous exosomes and thrombin in "signal-on" and "signal-off" modes, respectively, which are further applied to the analysis of complex biological samples successfully. Overall, the encapsulation of multi-enzyme with DFs proposes a promising strategy to regulate the microenvironment of biocatalytic cascades, which hold great potential in biotechnology, bioanalysis and disease diagnosis.

Ratiometric Dual Signal-Enhancing-Based Electrochemical Biosensor for Ultrasensitive Kanamycin Detection.

Based on the signal amplification elements of planar VS2/AuNPs nanocomposites and CoFe2O4 nanozyme, we herein developed an electrochemical biosensor for sensitive kanamycin (Kana) quantification. A ratiometric sensing platform was presented by incorporating VS2/AuNPs nanocomposites as a support material with excellent conductivity and high specific surface area, as well as hairpin DNA (hDNA) with complementary hybridization of biotinylated Kana-aptamer. In addition, streptavidin-functionalized CoFe2O4 nanozyme with superior peroxidase-like catalytic activity were immobilized onto the aptasensor, hence the peroxidase-like catalytic reaction could yield amplified electrochemical signals. With the presence of Kana, the aptamer-biorecognition resulted in a quantitative decrease of nanozyme accumulation and an increase of methylene blue response. Under optimal conditions, the electrochemical signal ratio of the aptasensor revealed a linear relation along with the logarithmic concentration of Kana from 1 pM to 1 µM, with the limit of detection reaching to 0.5 pM. Moreover, this aptasensor exhibited excellent precision, as well as high repeatability, hence possessing potentials in real samples and for diverse targets detection by easy replacement of the matched aptamer.

Gold nano-flowers (Au NFs) modified screen-printed carbon electrode electrochemical biosensor for label-free and quantitative detection of glycated hemoglobin.

Glycated hemoglobin (HbA1c) represents the average glucose level over the past three months and has been considered as the most important biomarker for the diagnosis of Type Ⅱ diabetes (T2D). Herein, a label-free and quantitative electrochemical biosensor based on 4-mercaptophenylboronic acid (4-MPBA) modified gold nano-flowers (Au NFs) substrate was developed for the determination of HbA1c. Under optimal conditions, the linear dynamic ranges of HbA1c (5 µg/mL - 1000 µg/mL) and HbA1c% (2%-20%) by cyclic voltammetry were achieved. The electrochemical biosensor showed great detection specificity towards HbA1c and relatively stability after storage at 4 °C. This method could also be applied in human serum system which holds great potential to be applied to monitor real blood samples of diabetes patients. In human serum system, the recovery rate could reach 103.8% and 99.0%. It could achieve fast detection, the total analysis time was less than 65 min, and the detection time was less than 10 s. Moreover, in terms of fabrication process, operation procedure, detection time and cost, this technique was superior to the current HbA1c detection methods suggesting great promise for the practical clinical use in the future.