Electrical graphene aptasensor for ultra-sensitive detection of anthrax toxin with amplified signal transduction.

Detection of the anthrax toxin, the protective antigen (PA), at the attomolar (aM) level is demonstrated by an electrical aptamer sensor based on a chemically derived graphene field-effect transistor (FET) platform. Higher affinity of the aptamer probes to PA in the aptamer-immobilized FET enables significant improvements in the limit of detection (LOD), dynamic range, and sensitivity compared to the antibody-immobilized FET. Transduction signal enhancement in the aptamer FET due to an increase in captured PA molecules results in a larger 30 mV/decade shift in the charge neutrality point (Vg,min ) as a sensitivity parameter, with the dynamic range of the PA concentration between 12 aM (LOD) and 120 fM. An additional signal enhancement is obtained by the secondary aptamer-conjugated gold nanoparticles (AuNPs-aptamer), which have a sandwich structure of aptamer/PA/aptamer-AuNPs, induce an increase in charge-doping in the graphene channel, resulting in a reduction of the LOD to 1.2 aM with a three-fold increase in the Vg,min shift.

Mogul-Patterned Elastomeric Substrate for Stretchable Electronics.

A mogul-patterned stretchable substrate with multidirectional stretchability and minimal fracture of layers under high stretching is fabricated by double photolithography and soft lithography. Au layers and a reduced graphene oxide chemiresistor on a mogul-patterned poly(dimethylsiloxane) substrate are stable and durable under various stretching conditions. The newly designed mogul-patterned stretchable substrate shows great promise for stretchable electronics.

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

Ultrarapid and ultrasensitive electrical detection of proteins in a three-dimensional biosensor with high capture efficiency.

The realization of a high-throughput biosensor platform with ultrarapid detection of biomolecular interactions and an ultralow limit of detection in the femtomolar (fM) range or below has been retarded due to sluggish binding kinetics caused by the scarcity of probe molecules on the nanostructures and/or limited mass transport. Here, as a new method for the highly efficient capture of biomolecules at extremely low concentration, we tested a three-dimensional (3D) platform of a bioelectronic field-effect transistor (bio-FET) with vertically aligned and highly dense one-dimensional (1D) ZnO nanorods (NRs) as a sensing surface capped by an ultrathin TiO2 layer for improved electrolytic stability on a chemical-vapor-deposited graphene (Gr) channel. The ultrarapid detection capability with a very fast response time (∼1 min) at the fM level of proteins in the proposed 3D bio-FET is primarily attributed to the fast binding kinetics of the probe-target proteins due to the small diffusion length of the target molecules to reach the sensor surface and the substantial number of probe molecules available on the largely increased surface area of the vertical ZnO NRs. This new 3D electrical biosensor platform can be easily extended to other electrochemical nanobiosensors and has great potential for practical applications in miniaturized biosensor integrated systems.

Toxicity of graphene nanoflakes evaluated by cell-based electrochemical impedance biosensing.

Graphene nanoflake toxicity was analyzed using cell-based electrochemical impedance biosensing with interdigitated indium tin oxide (ITO) electrodes installed in a custom-built mini-incubator positioned on an inverted optical microscope. Sensing with electrochemical measurements from interdigitated ITO electrodes was highly linear (R(2) = 0.93 and 0.96 for anodic peak current (Ipa) and cathodic peak current (Ipc), respectively). Size-dependent analysis of Graphene nanoflake toxicity was carried out in a mini-incubator system with cultured HeLa cells treated with Graphene nanoflakes having an average size of 80 or 30 nm for one day. Biological assays of cell proliferation and viability complemented electrochemical impedance measurements. The increased toxicity of smaller Graphene nanoflakes (30 nm) as measured by electrochemical impedance sensing and optical monitoring of treated cells was consistent with the biological assay results. Cell-based electrochemical impedance biosensing can be used to assess the toxicity of nanomaterials with different biomedical and environmental applications.

Reduction in oxidative stress during cellular responses to chemically functionalised graphene.

The two-dimensional nanocarbon material graphene (Gr) has been extensively studied due to its many potential biomedical applications including regenerative medicine, drug delivery, bioimaging, and biosensing. The effects of nitrogen-functionalisation on chemically driven Gr (CDG) cellular responses were studied by investigating the generation of reactive oxygen species (ROS) and mitochondrial morphology as well as focal adhesion, shape, proliferation and viability of HeLa cells grown on functionalised CDG (f-CDG) films. The drop casting of CDG nanosheets formed thin CDG films and the formation of nitrogen groups on the f-CDG thin films was mediated by N2 plasma treatment without the formation of observable surface defects. N-containing functional groups on the CDG thin films contributed to an increase in hydrophilicity. The proliferation and viability of HeLa cells grown on the f-CDG thin films were enhanced compared to those grown on CDG films alone and control samples. N-functionalisation of CDG thin films effectively reduced the ROS generated from cells on the f-CDG films. These results indicate that N2 plasma treatment of CDG is very useful in improving biocompatibility for the bio-application of graphene materials.

pH sensing characteristics and biosensing application of solution-gated reduced graphene oxide field-effect transistors.

Solution-gated reduced graphene oxide field-effect transistors (R-GO FETs) were investigated for pH sensing and biochemical sensing applications. A channel of a networked R-GO film formed by self-assembly was incorporated as a sensing layer into a solution-gated FET structure for pH sensing and the detection of acetylcholine (Ach), which is a neurotransmitter in the nerve system, through enzymatic reactions. The fabricated R-GO FET was sensitive to protons (H(+)) with a pH sensitivity of 29 mV/pH in terms of the shift of the charge neutrality point (CNP), which is attributed to changes in the surface potential caused by the interaction of protons with OH surface functional groups present on the R-GO surface. The R-GO FET immobilized with acetylcholinesterase (AchE) was used to detect Ach in the concentration range of 0.1-10mM by sensing protons generated during the enzymatic reactions. The results indicate that R-GO FETs provide the capability to detect protons, demonstrating their applicability as a biosensing device for enzymatic reactions.

Reduced graphene oxide field-effect transistor for label-free femtomolar protein detection.

We report reduced graphene oxide field effect transistor (R-GO FET) biosensor for label-free ultrasensitive detection of a prostate cancer biomarker, prostate specific antigen/α1-antichymotrypsin (PSA-ACT) complex. The R-GO channel in the device was formed by reduction of graphene oxide nanosheets networked by a self-assembly process. Immunoreaction of PSA-ACT complexes with PSA monoclonal antibodies on the R-GO channel surface caused a linear response in the shift of the gate voltage, V(g,min), where the minimum conductivity occurs. The R-GO FET can detect protein-protein interactions down to femtomolar level with a dynamic range over 6-orders of magnitude in the V(g,min) shift as a sensitivity parameter. High association constants of 3.2 nM(-1) and 4.2 nM(-1) were obtained for the pH 6.2 and pH 7.4 analyte solutions, respectively. The R-GO FET biosensor showed a high specificity to other cancer biomarker in the phosphate buffered saline solutions as well as in the human serum.