Graphene-based Aptasensor Research Articles

Laser-induced graphene-based aptasensor for the selective detection of Escherichia coli in urine samples

A Laser-Induced Graphene-based (LIG) electrode covalently functionalized with an aptamer (P12-55) was used to develop an aptasensor detecting Escherichia coli in urine samples. Recent strides in material science have spotlighted LIG for exceptional attributes like robust mechanical resistance, superior conductivity, extensive surface area, and facile synthesis/patterning on various polymeric substrates. Variations in the aptasensor charge transfer resistance upon interaction with bacterial cells were evaluated by electrochemical impedance spectroscopy. Tests in phosphate buffer saline solution showed the aptasensor linear response for E. coli between 100 and 103 CFU/mL. The sensor proved to be selective for E. coli as a negligible response was observed in the presence of Staphylococcus aureus or Pseudomonas aeruginosa. Finally, the sensor was calibrated in urine samples spiked with a known concentration of E. coli cells. Its characteristics make the aptasensor viable for low-cost and portable devices identifying pathogenic microorganisms at the point-of-need. Due to the short response time (about 30 min), we believe that these sensing devices may significantly improve control and management of urinary tract infections.

Aggregation of Apo/Glycated Human Serum Albumins and Aptamer-Saturated Graphene Quantum Dot: A Simulation Study.

Human serum albumin (HSA) is a protein carrier that transports a wide range of drugs and nutrients. The amount of glycated HSA (GHSA) is used as a diabetes biomarker. To quantify the GHSA amount, the fluorescent graphene-based aptasensor has been a successful method. In aptasensors, the key mechanism is the adsorption/desorption of albumin from the aptamer-graphene complex. Recently, the graphene quantum dot (GQD) has been reported to be an aptamer sorbent. Due to its comparable size to aptamers, it is attractive enough to explore the possibility of GQD as a part of an albumin aptasensor. Therefore, molecular dynamics (MD) simulations were performed here to reveal the binding mechanism of albumin to an aptamer-GQD complex in molecular detail. GQD saturated by albumin-selective aptamers (GQDA) is studied, and GHSA and HSA are studied in comparison to understand the effect of glycation. Fast and spontaneous albumin-GQDA binding was observed. While no specific GQDA-binding site on both albumins was found, the residues used for binding were confined to domains I and III for HSA and domains II and III for GHSA. Albumins were found to bind preferably to aptamers rather than to GQD. Lysines and arginines were the main contributors to binding. We also found the dissociation of GLC from all GHSA trajectories, which highlights the role of GQDA in interfering with the ligand binding affinity in Sudlow site I. The binding of GQDA appears to impair albumin structure and function. The insights obtained here will be useful for the future design of diabetes aptasensors.

Comparative Study of Field-Effect Transistors Based on Graphene Oxide and CVD Graphene in Highly Sensitive NT-proBNP Aptasensors.

Graphene-based materials are actively being investigated as sensing elements for the detection of different analytes. Both graphene grown by chemical vapor deposition (CVD) and graphene oxide (GO) produced by the modified Hummers' method are actively used in the development of biosensors. The production costs of CVD graphene- and GO-based sensors are similar; however, the question remains regarding the most efficient graphene-based material for the construction of point-of-care diagnostic devices. To this end, in this work, we compare CVD graphene aptasensors with the aptasensors based on reduced GO (rGO) for their capabilities in the detection of NT-proBNP, which serves as the gold standard biomarker for heart failure. Both types of aptasensors were developed using commercial gold interdigitated electrodes (IDEs) with either CVD graphene or GO formed on top as a channel of liquid-gated field-effect transistor (FET), yielding GFET and rGO-FET sensors, respectively. The functional properties of the two types of aptasensors were compared. Both demonstrate good dynamic range from 10 fg/mL to 100 pg/mL. The limit of detection for NT-proBNP in artificial saliva was 100 fg/mL and 1 pg/mL for rGO-FET- and GFET-based aptasensors, respectively. While CVD GFET demonstrates less variations in parameters, higher sensitivity was demonstrated by the rGO-FET due to its higher roughness and larger bandgap. The demonstrated low cost and scalability of technology for both types of graphene-based aptasensors may be applicable for the development of different graphene-based biosensors for rapid, stable, on-site, and highly sensitive detection of diverse biochemical markers.

Label-free electrochemical aptasensor for detection of Acinetobacter baumannii: Unveiling the kinetic behavior of reduced graphene oxide v/s graphene oxide

In this study, we introduce an electrochemical aptasensor for the sensitive detection of Acinetobacter baumannii. This method, based on impedance and current analysis, represents a first-of-its kind platform in the field of A. baumannii detection. We employed a screen-printed carbon electrode modified with both graphene oxide (GO) and reduced graphene oxide (rGO) as the immobilization platform, and the bacteria specific aptamer was employed as the biorecognition element. The aptamer selectively interacts with A. baumannii, causing a decrease in the peak current of the redox complex because the binding biomolecules on the electrode surface hinder the electron transfer of the redox probe. This initial point-of-care-compatible rGO electrochemical aptasensor demonstrated an exceptional sensing performance with an impressive limit of detection of 10 CFU/mL for detecting A. baumannii. This enhanced sensitivity can be attributed to several key factors including structural defects during reduction, increased surface area and improved electron transfer kinetics. The findings of this study highlight the potential of the developed aptasensor as a rapid and reliable detection platform for various pathogenic diseases. By further ensuring seamless integration into clinical practice, these label-free graphene-based aptasensors, with their π-π interactions and other favorable attributes, hold great promise for sensitive and specific pathogen detection, thereby contributing to advancements in healthcare diagnostics.

The binding of apo and glucose-bound human serum albumins to a free graphene sheet in aqueous environment: Simulation studies

Human serum albumin (HSA) is a blood protein serving as a carrier for a wide range of drugs and nutrients. A level of glycated HSA (GHSA) is used as a diabetes biomarker. A graphene-based aptasensor is one of potential techniques to detect GHSA. Not only the interactions of albumin and aptamer, but the albumin-graphene (GRA) binding mechanism are also crucial for developing a diabetes aptasensor. In this work, Molecular Dynamics simulations (MD) were employed to explore the binding of GRA to both GHSA and HSA. The GRA binding from the back and front sides of an albumin are fast and spontaneous. The multiple GRA binding sites are identified. GRA causes more denaturation of helical characteristics in GHSA (∼12% reduction of helical structure). Both back and front GRA adhesions generate comparable degrees of helical unfolding. Importantly, the presence of bound GRA induces the release of glucose from drug sites implying the loss of ligand-binding affinity. This loss of drug site activity is independent on the GRA binding positions because all bound positions lead to the exit of sugars. The GRA binding deconstructs not only secondary structure, but also albumin function. Apparently, GRA is a non-biocompatible material for albumin. To construct a potential graphene-based aptasensor to detect GHSA, it is necessary to be certain that no free GRA surface is available because a bare GRA can bind and denature both HSA and GHSA which can cause misleading data.

Novel Electrochemical Aptasensors Based on Bipolar Exfoliated Graphene for Label-Free Detection of Cancer Biomarkers

Early detection of cancer can noticeably increase the survival chance of many cancer patients. Quantifying cancer biomarkers detectable from blood is an efficient way for early detection of cancer diseases. Among various discovered cancer biomarkers, platelet-derived growth factor-BB (PDGF-BB) is an essential biomarker for early detection of cancer and monitoring cancer patients. This biomarker plays a vital role in developing and lymphatic metastasis of solid malignant tumors such as brain, lung, breast, and liver; which enlightens the importance of developing point-of-care (POC) biosensors for the detection of PDGF-BB. In recent years, the application of synthetic DNA or RNA-based bio-recognizers (i.e., aptamers) in cancer biomarker sensor development has been vastly investigated. Electrochemical label-free aptamer-based biosensors (also known as aptasensors) are highly suitable for POC. In this study, the application of bipolar exfoliated (BPE) graphene for developing PDGF-BB label-free aptasensors is investigated. Graphene as a single, two-dimensional layer of carbon atoms has very interesting features suitable for biosensor applications. The common graphene synthesis methods require complicated and costly processes and excessive use of harsh chemicals, as well as complex subsequent deposition procedures. In this study, a single setup has been used for exfoliation, reduction, and deposition of graphene nanosheets on a conductive electrode based on the principle of bipolar electrochemistry of graphite in deionized water. We investigated the properties of aptasensors based on graphene oxide (GO) deposited on a positive electrode feeding electrode and reduced-GO (rGO) deposited on a negative electrode feeding electrode. The PDGF-BB affinity aptamers were covalently immobilized by binding amino-tag terminated aptamers and carboxyl groups of GO and rGO surfaces. In this study, Fourier-transform infrared spectroscopy (FTIR) was used to study the surface characteristics of synthesized graphene and developed aptasensors. The scanning electron microscopy (SEM) was used to study the morphology of the bipolar exfoliated GO and rGO. The SEM study demonstrated that the rGO has a porous vertically aligned structure with pore sizes of around 100 nm, while GO has bulky flattened plates with random cracks. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used for characterizing the BPE graphene-based aptasensors in different stages of development and their sensing performances. The turn-off sensing strategy was implemented by measuring the peak-currents from DPV plots. The optimized aptasensor based on rGO showed a wide linear range of 0.75 pM-10 nM, high sensitivity of 7.83 A Logc-1 (unit of c, pM), and a low detection limit of 0.53 pM. However, the optimized aptasensor based on GO reached to its saturation point around 150 pM. This study demonstrated that bipolar electrochemistry is a simple yet efficient technique that could provide high-quality graphene for biosensing applications. Considering the BPE technique's simplicity and efficiency, this technique is highly promising for developing feasible and affordable lab-on-chip and point-of-care cancer diagnosis technologies.

A label-free electrochemical aptasensor for breast cancer cell detection based on a reduced graphene oxide-chitosan-gold nanoparticle composite

Regarding the cancer fatal consequences, early detection and progression monitoring are the most vital issues in patients’ treatment and mortality reduction. Therefore, there is a great demand for fast, inexpensive, and selective detection methods. Herein, a graphene-based aptasensor was designed for sensitive human breast cancer cell detection. A reduced graphene oxide-chitosan-gold nanoparticles composite was used as a biocompatible substrate for the receptor stabilization. The significant function of the aptamer on this composite is due to the synergistic effects of the components in improving the properties of the composite, including increasing the electrical conductivity and effective surface area. After the aptasensor incubation in MCF-7 cancer cells, the cell membrane proteins interacted specifically with the three dimensional-structure of the AS1411 aptamer, resulting in the cell capture on the aptasensor. The aptasensor fabrication steps were investigated by cyclic voltammetry and electrochemical impedance spectroscopy. The higher cell concentrations concluded to the higher captured cells on the aptasensor which blocked the Ferro/Ferricyanide access to the sensor, causing increases in the charge transfer resistances. This aptasensor shows a linear relationship with the cell concentration logarithm, high selectivity, a wide linear range of 1 × 101–1 × 106 cells/mL, and a low detection limit of 4 cells/mL.

Graphene-family materials in electrochemical aptasensors.

The study of graphene-based carbon nanocomposites has remarkably increased in recent years. Functionalized graphene-based nanostructures, including graphene oxide and reduced graphene oxide, have great potential as new innovative electrode materials in the fabrication of novel electrochemical sensors. Electrochemical sensors based on aptamers attracted great attention because of their high sensitivity and selectivity, and simple instrumentation, as well as low production cost. Aptamers as a potent alternative to antibodies are functional nucleic acids with a high tendency to specific analytes. Electrochemical aptasensors show specific recognition ability for a wide range of analytes. Although aptamers are selected in vitro in contrast to antibodies, they are interesting due to advantages like high stability, easy chemical modifications, and the potential to be employed in nanostructured device fabrication or electrochemical sensing devices. Recently, new nanomaterials have shown a significant impact on the production of electrochemical sensors with high efficiency and performance. This review aims to give an outline of electrochemical aptasensors based on the graphene family materials and discuss the detection mechanism in this type of aptasensors. The present review summarizes some of the recent achievements in graphene-based aptasensors and includes their recent electroanalytical applications. Graphical Abstract Graphical Abstract.

Graphene-based aptasensors: from molecule-interface interactions to sensor design and biomedical diagnostics.

Graphene-based nanomaterials have been widely utilized to fabricate various biosensors for environmental monitoring, food safety, and biomedical diagnostics. The combination of aptamers with graphene for creating biofunctional nanocomposites improved the sensitivity and selectivity of fabricated biosensors due to the unique molecular recognition and biocompatibility of aptamers. In this review, we highlight recent advances in the design, fabrication, and biomedical sensing application of graphene-based aptasensors within the last five years (2013-current). The typical studies on the biomedical fluorescence, colorimetric, electrochemical, electrochemiluminescence, photoelectrochemical, electronic, and force-based sensing of DNA, proteins, enzymes, small molecules, ions, and others are demonstrated and discussed in detail. More attention is paid to a few key points such as the conjugation of aptamers with graphene materials, the fabrication strategies of sensor architectures, and the importance of aptamers on improving the sensing performances. It is expected that this work will provide preliminary and useful guidance for readers to understand the fabrication of graphene-based biosensors and the corresponding sensing mechanisms in one way, and in another way will be helpful to develop novel high performance aptasensors for biological analysis and detection.

The Fabrication of High-Performance Flexible Mercury Aptasensor Based on Graphene

ABSTRACTLiquid-ion gated FET-type flexible graphene-based aptasensor was fabricated for Hg detection in real world samples. Single-layer graphene was grown and transferred onto a flexible substrate and integrated into the liquid-ion gated FET system via surface engineerging process. Field-induced responses to Hg2+ ions in real world samples were highly rapid, sensitive and selective, leading to the high-perfromance graphene aptasensor. The aptasensor also displayed excellent flexibility and mechanical durability.

High-Performance Flexible Graphene Aptasensor for Mercury Detection in Mussels

Mercury (Hg) is highly toxic but has been widely used for numerous domestic applications, including thermometers and batteries, for decades, which has led to fatal outcomes due to its accumulation in the human body. Although many types of mercury sensors have been developed to protect the users from Hg, few methodologies exist to analyze Hg(2+) ions in low concentrations in real world samples. Herein, we describe the fabrication and characterization of liquid-ion gated field-effect transistor (FET)-type flexible graphene aptasensor with high sensitivity and selectivity for Hg. The field-induced responses from the graphene aptasensor had excellent sensing performance, and Hg(2+) ions with very low concentration of 10 pM could be detected, which is 2-3 orders of magnitude more sensitive than previously reported mercury sensors using electrochemical systems. Moreover, the aptasensor showed a highly specific response to Hg(2+) ions in mixed solutions. The flexible graphene aptasensor showed a very rapid response, providing a signal in less than 1 s when the Hg(2+) ion concentration was altered. Specificity to Hg(2+) ions was demonstrated in real world samples (in this case samples derived from mussels). The aptasensor was fabricated by transferring chemical vapor deposition (CVD)-grown graphene onto a transparent flexible substrate, and the structure displayed excellent mechanical durability and flexiblility. This graphene-based aptasensor has potential for detecting Hg exposure in human and in the environment.

Biorecognition on Graphene: Physical, Covalent, and Affinity Immobilization Methods Exhibiting Dramatic Differences

The preparation of biorecognition layers on the surface of a sensing platform is a very crucial step for the development of sensitive and selective biosensors. Different protocols have been used thus far for the immobilization of biomolecules onto various electrode surfaces. In this work, we investigate how the protocol followed for the immobilization of a DNA aptamer affects the performance of the fabricated thrombin aptasensor. Specifically, the differences in selectivity and optimum amount of immobilized aptamer of the fabricated aptasensors adopting either physical, covalent, or affinity immobilization were compared. It was discovered that while all three methods of immobilization uniformly show a similar optimum amount of immobilized aptamer, physical, and covalent immobilization methods exhibit higher selectivity than affinity immobilization. Hence, it is believed that our findings are very important in order to optimize and improve the performance of graphene-based aptasensors.