Single-use disposable electrochemical label-free immunosensor for detection of glycated hemoglobin (HbA1c) using differential pulse voltammetry (DPV)

A single-use disposable in vitro electrochemical immunosensor for the detection of HbA1c in undiluted human serum using differential pulse voltammetry (DPV) was developed. A three-electrode configuration electrochemical biosensor consisted of 10-nm-thin gold film working and counter electrodes and a thick-film printed Ag/AgCl reference electrode was fabricated on a polyethylene terephthalate (PET) substrate. Micro-fabrication techniques including sputtering vapor deposition and thick-film printing were used to fabricate the biosensor. This was a roll-to-roll cost-effective manufacturing process making the single-use disposable in vitro HbA1c biosensor a reality. Self-assembled monolayers of 3-Mercaptopropionic acid (MPA) were employed to covalently immobilize anti-HbA1c on the surface of gold electrodes. Electrochemical impedance spectroscopy (EIS) and X-ray photoelectron spectroscopy (XPS) confirmed the excellent coverage of MPA-SAM and the upward orientation of carboxylic groups. The hindering effect of HbA1c on the ferricyanide/ferrocyanide electron transfer reaction was exploited as the HbA1c detection mechanism. The biosensor showed a linear range of 7.5-20 μg/mL of HbA1c in 0.1 M PBS. Using undiluted human serum as the test medium, the biosensor presented an excellent linear behavior (R2 = 0.999) in the range of 0.1-0.25 mg/mL of HbA1c. The potential application of this biosensor for in vitro measurement of HbA1c for diabetic management was demonstrated

A Rapidly Stabilizing Water-Gated Field-Effect Transistor Based on Printed Single-Walled Carbon Nanotubes for Biosensing Applications

Biosensors are expected to revolutionize disease management through provision of low-cost diagnostic platforms for molecular and pathogenic detection with high sensitivity and short response time. In this context, there has been an ever-increasing interest in using electrolyte-gated field-effect transistors (EG-FETs) for biosensing applications owing to their expanding potential of being employed for label-free detection of a broad range of biomarkers with high selectivity and sensitivity while operating at sub-volt working potentials. Although organic semiconductors have been widely utilized as the channel in EGFETs, primarily due to their compatibility with cost-effective low-temperature solution-processing fabrication techniques, alternative carbon-based platforms have the potential to provide similar advantages with improved electronic performances. Here, we propose the use of inkjet-printed polymer-wrapped monochiral singlewalled carbon nanotubes (s-SWCNTs) for the channel of EG-FETs in an aqueous environment. In particular, we show that our EG-CNTFETs require only an hour of stabilization before producing a highly stable response suitable for biosensing, with a drastic time reduction with respect to the most exploited organic semiconductor for biosensors. As a proof-of-principle, we successfully employed our water-gated device to detect the well-known biotin-streptavidin binding event

Direct recording of action potentials of cardiomyocytes through solution processed planar electrolyte-gated field-effect transistors

To achieve intracellular recording of action potentials by using simple devices that can be easily fabricated and processed is crucial in cardiology and neuroscience. Present tools and technology include invasive patch clamp technique, 3D nanostructures often combined with electro/opto poration methods and nanodevices such as nanowire field-effect transistors. However, these approaches mostly require complex manufacturing processes or are invasive. In this work, we report the spontaneous intracellular-like recording of cardiac cells using a cost-effective, planar Electrolyte-Gated Field-Effect Transistor (EGFET) based on solution-processed polymer-wrapped monochiral semiconducting single-walled carbon nanotubes (SWCNTs). By simply turning on the transistor, spontaneous recordings of intracellular-like action potentials of human induced pluripotent stem cells derived cardiomyocytes are enabled. In addition, we demonstrate that the same planar EGFET can also be employed as a platform for electroporation with significant device performance and cell viability. The simplicity of the device combined with the high signal to noise ratio opens up new opportunities for low-cost, reliable, and flexible biosensors and arrays for high quality parallel recording of cellular action potentials

A Single-Use, In Vitro Biosensor for the Detection of T-Tau Protein, A Biomarker of Neuro-Degenerative Disorders, in PBS and Human Serum Using Differential Pulse Voltammetry (DPV)

A single-use, in vitro biosensor for the detection of T-Tau protein in phosphate-buffer saline (PBS) and undiluted human serum was designed, manufactured, and tested. Differential pulse voltammetry (DPV) served as the transduction mechanism. This biosensor consisted of three electrodes: working, counter, and reference electrodes fabricated on a PET sheet. Both working and counter electrodes were thin gold film, 10 nm in thickness. Laser ablation technique was used to define the size and structure of the biosensor. The biosensor was produced using cost-effective roll-to-roll process. Self-assembled monolayers (SAM) of 3-mercaptopropionic acid (MPA) were employed to covalently immobilize the anti-T-Tau (T-Tau antibody) on the gold working electrode. A carbodiimide conjugation approach using N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) and N–hydroxysuccinimide (NHS) cross-linked anti-T-Tau to the carboxylic groups on one end of the MPA. A T-Tau protein ladder with six isoforms was used in this study. The anti-T-Tau concentration used was 500,000 pg/mL. The T-Tau protein concentration ranged from 1000 pg/mL to 100,000 pg/mL. DPV measurements showed excellent responses, with a good calibration curve. Thus, a practical tool for simple detection of T-Tau protein, a biomarker of neuro-degenerative disorders, has been successfully developed. This tool could also be extended to detect other biomarkers for neuro-degenerative disorders, such as P-Tau protein and β-amyloid 42

In Vitro Quantified Determination of β-Amyloid 42 Peptides, a Biomarker of Neuro-Degenerative Disorders, in PBS and Human Serum Using a Simple, Cost-Effective Thin Gold Film Biosensor

A simple in vitro biosensor for the detection of β-amyloid 42 in phosphate-buffered saline (PBS) and undiluted human serum was fabricated and tested based on our platform sensor technology. The bio-recognition mechanism of this biosensor was based on the effect of the interaction between antibody and antigen of β-amyloid 42 to the redox couple probe of K4Fe(CN)6 and K3Fe(CN)6. Differential pulse voltammetry (DPV) served as the transduction mechanism measuring the current output derived from the redox coupling reaction. The biosensor was a three-electrode electrochemical system, and the working and counter electrodes were 50 nm thin gold film deposited by a sputtering technique. The reference electrode was a thick-film printed Ag/AgCl electrode. Laser ablation technique was used to define the size and structure of the biosensor. Cost-effective roll-to-roll manufacturing process was employed in the fabrication of the biosensor, making it simple and relatively inexpensive. Self-assembled monolayers (SAM) of 3-Mercaptopropionic acid (MPA) was employed to covalently immobilize the thiol group on the gold working electrode. A carbodiimide conjugation approach using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) and N–hydroxysuccinimide (NHS) was undertaken for cross-linking antibody of β-amyloid 42 to the carboxylic groups on one end of the MPA. The antibody concentration of β-amyloid 42 used was 18.75 µg/mL. The concentration range of β-amyloid 42 in this study was from 0.0675 µg/mL to 0.5 µg/mL for both PBS and undiluted human serum. DPV measurements showed excellent response in this antigen concentration range. Interference study of this biosensor was carried out in the presence of Tau protein antigen. Excellent specificity of this β-amyloid 42 biosensor was demonstrated without interference from other species, such as T-tau protein

Single-use nonenzymatic glucose biosensor based on CuO nanoparticles ink printed on thin film gold electrode by micro-plotter technology

We reported a single-use non-enzymatic glucose biosensor based on a printed CuO nanoparticles film on thin gold film electrode. This three-electrode configuration biosensor consisted of a thin gold film (10 nm) working and counter electrodes and a thick-film printed Ag/AgCl reference electrode. The biosensor was fabricated on a polyethylene terephthalate (PET) substrate. CuO nanoparticles with an average particle size of ~ 7 nm were synthesized by a one-step precipitation approach, formulating an aqueous-based nanoparticles ink. A CuO nanoparticles square area of 0.9 × 0.9 mm2 was printed on thin film Au working electrode (φ = 1.4 mm) of the biosensor by micro-plotter technology. The remarkable resolution and dimensional accuracy of micro-plotter printing technology resulted in a better development of nano-catalyst biosensors compared to conventional ink-jet printing or other techniques. X-ray Photoelectron Spectroscopy (XPS) and Transmission Electron Microscopy (TEM) were used to characterize the printed biosensor. Field Emission Scanning Electron Microscopy (FESEM) characterizations of the copper oxide printed features showed a nano-porous morphology together with high printing resolution. Cyclic Voltammetry (CV) and chronoamperometry (CA) measurements revealed the considerable electro-catalytic performance of the printed biosensor having high sensitivity (2419.8 μA mM− 1 cm− 2), acceptable stability and excellent interference rejection against ascorbic acid (AA), uric acid (UA), dopamine (DA), lactose, mannose and maltose. Interference rejection studies were performed in undiluted human serum to verify the functionality of the sensor for real samples. The developed biosensor showed a linear response toward glucose in the range of 0.1 to 6.5 mM. The lower detection limit of the biosensor was measured as 0.5 μM glucose. This study suggested a high performance robust glucose biosensor not only suitable for biomedical single-use in-vitro application but also for long-term glucose monitoring in industrial processes