An ultrasensitive electrochemical aptasensor based on zeolitic imidazolate framework-67 loading gold nanoparticles and horseradish peroxidase for detection of aflatoxin B1.

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins and poses a high risk to human health. Highly sensitive and rapid detection is one of the most effective preventive measures to avoid potential hazards. Herein, an electrochemical aptasensor based on DNA nanotetrahedron and zeolitic imidazolate framework-67 loading gold nanoparticles, horseradish peroxidase, and aptamers was designed for the ultrasensitive detection of AFB1. The high specific surface area and large pore volume of zeolitic imidazolate framework-67 can increase the loading capacity and further improve the detection sensitivity of electrochemical aptasensors. DNA nanotetrahedron can enhance the capture ability of AFB1 with steady immobilization. The developed aptasensor showed good analytical performance for AFB1 detection, with a detection limit of 3.9 pg mL-1 and a wide linear range of 0.01-100 ng mL-1. The aptasensor detected AFB1 in corn samples with recovery rates ranging from 94.19%-105.77% and has potential for use in food safety monitoring.

Freestanding Nanofiber-Assembled Aptasensor for Precisely and Ultrafast Electrochemical Detection of Alzheimer's Disease Biomarkers.

Amyloid beta-protein (AßAß) is a main hallmark of Alzheimer's disease (AD), and a low amount of Aß protein accumulation appears to be a potential marker for AD. Here, an electrochemical DNA biosensor based on polyamide/polyaniline carbon nanotubes (PA/PANI-CNTs) is developed with the aim of diagnosing AD early using a simple, low-cost, and accessible method to rapidly detect Aß42 in human blood. Electrospun PA nanofibers served as the skeleton for the successive in situ deposition of PANI and CNTs, which contribute both high conductivity and abundant binding sites for the Aß42 aptamers. After the aptamers are immobilized, this aptasensor exhibits precise and specific detection of Aß42 in human blood within only 4 min with an extremely fast response rate, lower detection limit, and excellent linear detection range. These findings make a significant contribution to advancing the development of serum-based detection techniques for Aß42, thereby paving the way for improved diagnostic capabilities in the field of AD.

Enhanced Electrochemical Characterization of the Immune Checkpoint Protein PD-L1 using Aptamer-Functionalized Magnetic Metal-Organic Frameworks.

Programmed death ligand 1 (PD-L1) is highly expressed in cancer cells and participates in the immune escape process of tumor cells. However, as one of the most promising biomarkers for cancer immunotherapy monitoring, the key problem ahead of practical usage is how to effectively improve the detection sensitivity of PD-L1. Herein, an electrochemical aptasensor for the evaluation of tumor immunotherapy is developed based on the immune checkpoint protein PD-L1. The fundamental principle of this method involves the utilization of DNA nanotetrahedron (NTH)-based capture probes and aptamer-modified magnetic metal-organic framework nanocomposites as signaling probes. A synergistic enhancement is observed in the electrocatalytic effect between Fe3O4 and UiO-66 porous shells in Fe3O4@UiO-66 nanocomposites. Therefore, the integration of aptamer-modified Fe3O4@UiO-66@Au with NTH-assisted target immobilization as an electrochemical sensing platform can significantly enhance sensitivity and specificity for target detection. This method enables the detection of targets at concentrations as low as 7.76 pg mL-1 over a wide linear range (0.01 to 1000 ng mL-1). The authors have successfully employed this sensor for in situ characterization of PD-L1 on the cell surface and for monitoring changes in PD-L1 expression during drug therapy, providing a cost-effective yet robust alternative to highly expensive and expertise-dependent flow cytometry.

Integrating porphyrin-based nanoporous organic polymers with electrochemical aptasensors for ultratrace detection of kanamycin.

The synthesis and characterization of two new porphyrin-based porous organic polymers (POPs) via Sonogashira cross-coupling reaction and leverage the two obtained POPs is reported for the fabrication of electrochemical aptasensors to detect kanamycin at an ultratrace level. The resultant electrochemical aptasensor demonstrates a high linear relationship with the logarithmic value of kanamycin concentration in the range 5 × 10-5-5 µg/L with the limit of detection of 17.6 pg/L or 36.3 fM. During the analysis of real samples from milk and river, a relative standard deviation of less than 4.39%, and good recovery values in the range 97.0-105% were obtained.

Microneedle electrochemical aptamer-based sensing: Real-time small molecule measurements using sensor-embedded, commercially-available stainless steel microneedles.

Microneedle sensors could enable minimally-invasive, continuous molecular monitoring - informing on disease status and treatment in real-time. Wearable sensors for pharmaceuticals, for example, would create opportunities for treatments personalized to individual pharmacokinetics. Here, we demonstrate a commercial-off-the-shelf (COTS) approach for microneedle sensing using an electrochemical aptamer-based sensor that detects the high-toxicity antibiotic, vancomycin. Wearable monitoring of vancomycin could improve patient care by allowing targeted drug dosing within its narrow clinical window of safety and efficacy. To produce sensors, we miniaturize the electrochemical aptamer-based sensors to a microelectrode format, and embed them within stainless steel microneedles (sourced from commercial insulin pen needles). The microneedle sensors achieve quantitative measurements in body-temperature undiluted blood. Further, the sensors effectively maintain electrochemical signal within porcine skin. This COTS approach requires no cleanroom fabrication or specialized equipment, and produces individually-addressable, sterilizable microneedle sensors capable of easily penetrating the skin. In the future, this approach could be adapted for multiplexed detection, enabling real-time monitoring of a range of biomarkers.

Bioanalytical methods encompassing label-free and labeled tuberculosis aptasensors: A review.

The high incidence of tuberculosis (TB) infection is of great concern world-wide. The traditional TB diagnostic techniques are not ideal for TB diagnosis in resource-poor countries. This is due to their high complexity, expensive nature, long time duration, poor sensitivity and specificity, as well as their requirement for sophisticated laboratories with special biosafety conditions. These limitations are major factors contributing to late diagnosis of TB and its continuous persistence. Biosensors offer several advantages as diagnostic tool due to their independence on Mycobacterium tuberculosis, simplicity, low-cost, high sensitivity and specificity. Likewise, the increasing interest on aptamers as biorecognition elements in biosensor application lies in their ease of synthesis and modification, chemical and thermal stability, low-cost, and high sensitivity and specificity towards their targets. Several research works have been done and many more are still on-going on fabrication and application of aptasensors for TB diagnosis. This review summarizes the label-free and label-based electrochemical, piezoelectric and optical aptasensors for TB diagnosis published in the last decade. It focuses on their various aptamer sequence modifications, assay formats, sensitivities, nanomaterial and enzyme-based signal amplification strategies, and the possibility of miniaturization and automation using microfluidic devices.

Recent advances in nanomaterials-based optical and electrochemical aptasensors for detection of cyanotoxins.

The existence of cyanotoxins poses serious threats to human health, it is highly desirable to develop specific and sensitive methods for rapid detection of cyanotoxins in food and water. Due to the distinct advantages of aptamer including high specificity, good stability and easy preparation, various aptamer-based sensors (aptasensors) have been proposed to promote the detection of cyanotoxins. In this review, we summarize recent advance in optical and electrochemical aptasensors for cyanotoxins sensing by integrating with versatile nanomaterials or innovative sensing strategies, such as colorimetric aptasensors, fluorescent aptasensors, surface enhancement Raman spectroscopy-based aptasensors, voltammetric aptasensors, electrochemical impedance spectroscopy-based aptasensors and photoelectrochemical aptasensors. We highlight the accomplishments and advancements of aptasensors with improved performance. Furthermore, the current challenges and future prospects in cyanotoxins detection are discussed from our perspectives, which we hope to provide more ideas for future researchers.

An electrochemical aptasensor based on intelligent walking DNA nanomachine with cascade signal amplification powered by nuclease for Mucin 1 assay.

Intelligently walking DNA nanomachines have sprung up an upsurge in various nucleic acid testing, but the rapid and sensitive test methods toward disease biomarker proteins based on the signal amplification strategy of DNA nanomachines were still ongoing development. In this work, an electrochemical aptasensor coupling the magnetic separation technique with the nuclease-powered walking DNA nanomachine was established for Mucin 1 (MUC1) detection. The magnetic beads (MBs) were modified by MUC1 aptamer hybridized with blocker DNA probe (BDP). After reacting with MUC1 proteins, the BDP was released from MBs to trigger the opening of capture hairpin DNA on Au nanoparticle (Au NPs)/MXene-modified electrode surface. In the presence of exonuclease III (Exo III), the BDP as a DNA walker is activated to autonomously move on the electrode. Then, lots of residual DNA fragments can still stay on electrode, further hybridizing with hairpin DNA, which can capture more UiO-66-NH2 metal-organic frameworks (MOFs). The amounts of ligands in MOFs can generate enhanced differential pulse voltammetry (DPV) signal probes. Furthermore, the concentrations of MUC1 can convert into the amplified DPV signals by introducing the signal amplification between the BDP as DNA walkers and Exo III as driven forces. This proposed electrochemical aptasensor achieved MUC1 detection ranging from 5 pg/mL to 50 ng/mL with detection limit of 0.72 pg/mL. Consequently, the designed and nuclease-powered walking DNA nanomachine provided an efficient strategy for the quantitative analysis of proteins by the interconversion between protein and BDP as a walker, which exhibited practical applicability of MUC1 detection in human serum.

Electrochemical Aptasensors for Parkinson's Disease Biomarkers Detection.

BACKGROUND: Biomarkers are characteristic molecules that can serve as indicators of biological process status or condition; here, they are being studied with special relevance to Parkinson's Disease (PD). This disease is a chronic neurodegenerative disorder very difficult to study given the site of pathology and due to a clinical phenotype that fluctuates over time. Currently, there is no definitive diagnostic test for Parkinson's Disease; thus, clinicians hope that the detection of crucial biomarkers will help in the symptomatic and presymptomatic diagnostics and provide surrogate endpoints to demonstrate the clinical efficacy of new treatments. METHODS: Electrochemical aptasensors are excellent analytical tools that are used in the detection of PD biomarkers, as they are portable, easy to use, and perform real-time analysis. RESULTS: In this review, we discuss the most important clinical biomarkers for PD, highlighting their physiological role and function in the disease. Herein, we review, for the first time, innovative aptasensors for the detection of current potential PD biomarkers based on electrochemical techniques and discuss future alternatives, including ideal analytical platforms for point-of-care diagnostics. CONCLUSION: These new tools will be critical not only in the discovery of sensitive, specific, and reliable biomarkers of preclinical PD, but also in the development of tests that can assist in the early detection and differential diagnosis of parkinsonian disorders and in monitoring disease progression. Various methods for fixing aptamers onto the sensor surfaces, enabling quantitative and specific PD biomarker detection present in synthetic and clinical samples, will also be discussed.

Ultrasensitive Detection of Ochratoxin A With a Zeolite Imidazolate Frameworks Composite-Based Electrochemical Aptasensor.

Ochratoxin A (OTA) is a harmful mycotoxin, which is mainly secreted by Penicillium and Aspergillus species. In this work, an electrochemical aptasensor is presented for OTA detection based on Au nanoparticles (AuNPs) modified zeolite imidazolate frameworks (ZIFs) ZIF-8 platform and duplex-specific nuclease (DSN) triggered hybridization chain reaction (HCR) signal amplification. G-quadruplex-hemin assembled HCR nanowire acted as a nicotinamide adenine dinucleotide (NADH) oxidase and an HRP-mimicking DNAzyme. Besides, thionine (Thi) was enriched as a redox probe for signal amplification in this pseudobienzyme electrocatalytic system. Under the optimal conditions, the analytical response ranged from 1 to 107 fg ml-1 with a detection limit of 0.247 fg ml-1. Furthermore, the aptasensor was proven to be applied in real wheat samples with a recovery between 96.8 and 104.2%, illustrating the potential prospects in practical detection.

Smartphone assisted portable biochip for non-invasive simultaneous monitoring of glucose and insulin towards precise diagnosis of prediabetes/diabetes.

Diabetes is a chronic disease with metabolic disorders. Prediabetes is a condition in which the blood glucose levels are not high enough to be diagnosed as diabetes, but insulin resistance and ß-cell dysfunction are simultaneously present. Glucose and insulin are important biomarkers for diagnosing diabetes/prediabetes. It is critical and essential to monitor glucose and insulin levels to achieve the early and precise diagnosis, timely prevention and scientific management of diabetes/prediabetes. Here, an electrochemical aptasensor on screen-printed carbon electrode (SPCE) was developed for non-invasive simultaneous real-time detection of glucose and insulin in saliva. Two thiolated aptamers specific to glucose and insulin, respectively, were fabricated onto gold nanoparticles (AuNPs) decorated SPCE through Au-S bonds to form the sensing interface terminated with redox probes methylene blue (SPCE-AuNPs-GluApt-MB and SPCE-AuNPs-InsApt-MB). With the electrochemical signal readout on SPCE, the sensing interface responded to glucose linearly in the range of 0.1-50 mM with a detection limit of 0.08 mM, and responded linearly to insulin in the range of 0.05-15 nM with a detection limit of 0.85 nM. Integration of a SPCE based sensing interface with a portable wireless biochip, the continuous real-time detection of glucose and insulin in saliva was successfully realized with smartphone signal readout in a non-invasive way, demonstrating its potential as a non-invasive and reliable tool for precise diagnosis of diabetes/prediabetes in a point of care fashion.

Conjugated bimetallic cobalt/iron polyphthalocyanine as an electrochemical aptasensing platform for impedimetric determination of enrofloxacin in diverse environments.

The synthesis of bimetallic cobalt/iron polyphthalocyanine (represented by polyCoFePc) network via a modified solid-phase synthesis method is described. It was exploited as a platform for anchoring enrofloxacin (ENR)-targeted aptamer strands, thus, fabricating a label-free impedimetric aptasensor for determination of ENR. The polyCoFePc exhibited a porous two-dimensional (2D) conjugated nanostructure and rich functional groups, and showed a superior binding interaction toward aptamer strands as compared to monometallic polyFePc and polyCoPc networks. This finding was attributed to structural defects and increased active binding sites, thereby giving a highly sensitive detection ability toward ENR. By using electrochemical impedance spectroscopy (EIS), the polyCoFePc-based electrochemical aptasensor exhibited an extremely low detection limit of 0.06 fg mL-1 within the ENR concentration from 0.1 fg mL-1 to 100 pg mL-1, along with high selectivity, good reproducibility, and remarkable stability. Interestingly, the constructed polyCoFePc-based aptasensor also demonstrated wide practicability in various environments. The recoveries of ENR spiked into river water, milk, and pork samples ranged within 91.2 - 107.2%, 90.5 - 109.6%, and 91.2 - 102.3%, respectively.

Electrochemical aptamer-based sensors for food and water analysis: A review.

Global food and water safety issues have prompted the development of highly sensitive, specific, and fast analytical techniques for food and water analysis. The electrochemical aptamer-based detection platform (E-aptasensor) is one of the more promising detection techniques because of its unique combination of advantages that renders these sensors ideal for detection of a wide range of target analytes. Recent research results have further demonstrated that this technique has potential for real world analysis of food and water contaminants. This review summaries the recently developed E-aptasensors for detection of analytes related to food and water safety, including bacteria, mycotoxins, algal toxins, viruses, drugs, pesticides, and metal ions. Ten different electroanalytical techniques and one opto-electroanalytical technique commonly employed with these sensors are also described. In addition to highlighting several novel sensor designs, this review also describes the strengths, limitations, and current challenges this technology faces, and future development trend.

Electrochemical aptasensor for multi-antibiotics detection based on endonuclease and exonuclease assisted dual recycling amplification strategy.

An ultrasensitive electrochemical aptasensor for multiplex antibiotics detection based on endonuclease and exonuclease assisted dual recycling amplification strategy was proposed. Kanamycin and chloramphenicol were selected as candidates. Firstly, aptamers of the antibiotics were immobilized on bar A and then binding with their endonuclease labeled complementary DNA strands to construct enzyme-cleavage probes. Secondly, The nano zirconium-metal organic framework (NMOF) particles with 1,4-benzene-dicarboxylate (BDC) as linker was defined as UiO-66. And its updated version, hierarchically porous UiO-66 (HP-UIO-66) decorated with different electroactive materials as signal tags were synthesized. Then they were immobilized on bar B linked by two duplex DNA strands which can be specifically cleaved by corresponding enzyme-cleavage probes in bar A. Once targets were introduced into system, aptamers can capture them and then release enzyme-cleavage probes. In the presence of exonuclease-I, exonuclease assisted target recycling amplification was triggered and more enzyme-cleavage probes were released into solution. The probes can trigger endonuclease assisted recycles and repeatedly cleave their corresponding duplex DNA strands on bar B then released numerous signal tags into supernatant. Thus two recycling amplification was performed in the system. Finally, MB and Fc in the signal tags were detected by square wave voltammetry after removing bar A/B and the current intensities were correspondent with the concentration of KANA and CAP respectively. Under the optimum condition, the limits of detection for the KANA and CAP were 35fM and 21fM respectively with a wide linear range from 1 × 10-4 to 50nM. This dual recycling amplification detection system exhibited high sensitivities and specificity. It can be easily extended to detect other targets if changing the corresponding aptamers and has potential application values for screening of multiplex antibiotics residues in food safety.

Thrombin aptasensor enabled by Pt nanoparticles-functionalized Co-based metal organic frameworks assisted electrochemical signal amplification.

In this work, a Pt nanoparticles-functionalized Co-based metal organic frameworks (PtNPs@Co(II)MOFs@PtNPs) was synthesized and applied in electrochemical aptasensor for thrombin (TB) detection. First, the Co(II)MOFs@PtNPs were prepared via the mixed solvothermal method, which consists of inner Pt nanoparticles (PtNPs) encapsulated by aminofunctionalized Co(II)MOFs materials. Following that, additional PtNPs were adsorbed on the surface of Co(II)MOFs@PtNPs, resulting in the formation of PtNPs@Co(II)MOFs@PtNPs nanocomposite. The PtNPs@Co(II)MOFs@PtNPs nanocomposites with a large surface area were implimented as nanocarriers to immobilize a mass of TBA II for the formation of the TBA II bioconjugates that could be captured onto the electrode surface by sandwich-type format. Moreover, the PtNPs@Co(II)MOFs@PtNPs nanocomposites could directly use as redox tags for charge-generating and electron-transporting with the electron transfer from Co(II) to Co(III). Furthermore, in the presence of H2O2, the PtNPs@Co(II)MOF@PtNPs could effectively catalyze H2O2 oxidation with improvement electron transfer of redox probe, resulting in electrochemical signal amplification. Based on the above superior advantages, TB was determined in the concentration range from 0.1pM to 50nM with a detection limit of 0.33fM. Furthermore, the excellent sensitivity and selectivity can be easily established for quantitative analysis of other analytes.

Carbon nanomaterials-based electrochemical aptasensors.

Carbon nanomaterials (CNMs) have attracted increasing attention due to their unique electrical, optical, thermal, mechanical and chemical properties. CNMs are extensively applied in electronic, optoelectronic, photovoltaic and sensing devices fields, especially in bioassay technology. These excellent properties significantly depend on not only the functional atomic structures of CNMs, but also the interactions with other materials, such as gold nanoparticles, SiO2, chitosan, etc. This review systematically summarizes applications of CNMs in electrochemical aptasensors (ECASs). Firstly, definition and development of ECASs are introduced. Secondly, different ways of ECASs about working principles, classification and construction of CNMs are illustrated. Thirdly, the applications of different CNMs used in ECASs are discussed. In this review, different types of CNMs are involved such as carbon nanotubes, graphene, graphene oxide, etc. Besides, the newly emerging CNMs and CNMs-based composites are also discoursed. Finally, we demonstrate the future prospects of CNMs-based ECASs, and some suggestions about the near future development of CNMs-based ECASs are highlighted.